Mitiglinide

• MF C₁₉H₂₅NO₃
• MW 315.407 Da

Mitiglinide (INN, trade name Glufast) is a drug for the treatment of type 2 diabetes.^[1]

Mitiglinide belongs to the meglitinide class of blood glucose-lowering drugs and is currently co-marketed in Japan by Kissei and Takeda. The North America rights to mitiglinide are held by Elixir Pharmaceuticals. Mitiglinide has not yet gained FDA approval.

Pharmacology

Mitiglinide is thought to stimulate insulin secretion by closing the ATP-sensitive K(+) K(ATP) channels in pancreatic beta-cells.

Dosage

Mitiglinide is delivered in tablet form.

Mitiglinide calcium hydrate

The condensation of dimethyl succinate (I) with benzaldehyde (II) by means of NaOMe in refluxing methanol followed by hydrolysis with NaOH in methanol/water gives 2-benzylidenesuccinic acid (III). Compound (III) is treated with refluxing Ac2O, yielding the corresponding anhydride (IV), which by reaction with cis-perhydroisoindole (V) in toluene affords the monoamide (VI). This amide is reduced with H2 over a chiral Rhodium catalyst and treated with (R)-1-phenylethylamine (VII) to provide the chiral salt (VIII) as a single diastereomer isolated by crystallization. Finally, this salt is treated first with aqueous NH4OH and then with aqueous CaCl2.

he optical resolution of racemic 2-benzylsuccinic acid (XV) using the chiral amines (R)-1-phenylethylamine (VII), (R)-1-(1-naphthyl)ethylamine (XIV) or (S)-1-phenyl-2-(4-tolyl)ethylamine (XVI) is carried out by fractional crystallization of the corresponding diastereomeric salts and treatment with 2N HCl, providing the desired enantiomer 2(S)-benzylsuccinic acid (XVII). Reaction of (XVII) with SOCl2 gives the corresponding acyl chloride (XVIII), which is treated with 4-nitrophenol (XIX) and TEA in dichloromethane to yield the activated diester (XX). The regioselective reaction of (XX) with cis-perhydroisoindole (V) in dichloromethane affords the monoamide (XXI), which by reaction with HCl and methanol provides the corresponding methyl ester (XXII). This ester is hydrolyzed with NaOH to the previously described chiral succinamic acid (XIII), which is finally converted into its calcium salt.

PATENT

https://www.google.com/patents/WO2009047797A2?cl=en

Perhydroisoindole derivative, (S)-mitiglinide of formula I is a potassium channel antagonist for the treatment of type 2 diabetes mellitus and is chemically known as (5)-2-benzyl-3-(cis-hexahydro-2- isoindolinylcarbonyl) propionic acid.

Formula I

It has potent oral hypoglycemic activity and is structurally different from the sulphonylureas, although it stimulates calcium influx by binding to the sulphonylurea receptor on pancreatic β-cells and closing K⁺ _ATP channels. Perhydroisoindole derivatives including (S)-mitiglinide and salts thereof were first disclosed in US patent 5,202,335. This patent discloses preparation of (S)-mitiglinide by the reaction of (5)-3-benzyloxycarbonyl-4-phenylbutyric acid with cis-hexahydroisoindoline in the presence of N- methylmorpholine and isobutyl chloroformate followed by debenzylation with palladium on carbon in ethyl acetate to yield (5)-mitiglinide as viscous oil. (S)-Mitiglinide is isolated as its hemi calcium salt using calcium chloride in water which is further recrystallized with diisopropyl ether. Melting point of calcium salt of mitiglinide calcium dihydrate salt is herein reported as 179-185 ⁰C. (S)-Mitiglinide prepared by the above process is obtained in low yields. Further, the synthetic method described in the patent does not enable the desired regioselectivity. Extensive purification steps are required to obtain the desired compound, which makes the process unattractive from industrial point of view. US patent 6,133,454 discloses a process for the preparation of (S)-mitiglinide by reacting dimethyl succinate with benzaldehyde in methanolic medium, to yield a diacid which is converted to corresponding anhydride and is further reacted with the perhydroisoindole to yield 2-[(cis- perhydroisomdol^-ytycarbonylmethyl^-phenylacrylic acid which is then subjected to catalytic hydrogenation using the complex rhodium/(2S,4S)-N-butoxycarbonyl-4-diphenylphosphino-2-diphenyl- phosphino-methylpyrrolidine (Rh/(S,S) BPPM) as asymmetric hydrogenation catalyst, followed by conversion to pharmaceutically acceptable salt of (S)-mitiglinide. The above patent utilizes ruthenium complex which is expensive, carcinogenic and toxicity, hence not recommended for industrial scale. European patent publication no. EP 0967204 discloses the preparation of mitiglinide by deprotecting benzyl-(S)-2-benzyl-3-(cis-hexahydro-2-isoindolinyl-carbonyl) propionate and converting the same to calcium dihydrate salt in crystalline form using calcium chloride, water and ethanol. The crystals of calcium salt are further recrystallized using ethanol and water. But the patent is silent about the crystalline form of mitiglinide calcium.

It will be appreciated by those skilled in the art that perhydroisoindole derivative, (S)-mitiglinide of formula I contains a chiral centre and therefore exists as enantiomers. Optically active compounds have increasingly gained importance since the technologies to develop optically active compounds in high purity have considerably improved. Obtaining asymmetric molecules has traditionally involved resolving the desired molecule from a racemic mixture using a chiral reagent, which is not profitable as it increases the cost and processing time. Alternatively, desired enantiomer can be obtained by selective recrystallization of one enantiomer. However such a process is considered inefficient, in that product recovery is often low, purity is uncertain and more than 50% of the material is lost. Enantiomers can also be resolved chromatographically, although the large amount of solvent required for conventional batch chromatography is cost prohibitive and results in the preparation of relatively dilute products. Limited throughput volumes also often make batch chromatography impractical for large-scale production. Even so, it is a common experience for those skilled in the art to find chiral separation of certain chiral mixtures to be inefficient or ineffective, thereby resulting in the efforts towards development of newer methodologies for asymmetric synthesis.

It would be of significant advantage to obtain (.S)-mitiglinide by development of reaction conditions necessary for productive manufacture of the required (5)-enantiomer, substantially free of the unwanted (R)-enantiomer, in large quantities that meet acceptable pharmaceutical standards. It is the property of the solid compounds to exist in different polymorphic form. By the term polymorphs mean to include different physical forms, crystal forms, crystalline/liquid crystalline/non-crystalline (amorphous) forms. This has especially become very interesting after observing that many antibiotics, antibacterials, tranquilizers etc, exhibit polymorphism and some/one of the polymorphic forms of a given drug exhibit superior bio-availability and consequently show much higher activity compared to other polymorphs. It has also been disclosed that the amorphous forms in a number of drugs exhibit different dissolution characteristics and in some cases different bioavailability patterns compared to the crystalline form [Konne T., Chem. Pharm. Bull. 38, 2003 (1990)]. The solubility of a material is also influenced by its solid-state properties, and it has been suggested that the solubility of an amorphous compound is 10 to 1600 times higher than that of its most stable crystalline structures (Bruno C. Hancock and Michael Parks, ‘What is the true solubility advantage for amorphous pharmaceuticals’, Pharmaceutical Research 2000, Apr; 17(4):397-404). Thus it can be concluded that amorphous products are in general more soluble and often show improved absorption in humans.

Thus, there is a widely recognized need for developing a stable polymorph, which would further offer advantages over crystalline forms in terms of better dissolution and the availability profiles. Also none of the prior art references disclose amorphous form of mitiglinide calcium. Thus present invention provides amorphous form of mitiglinide calcium.

It is also required that the final API like mitiglinide whether in the amorphous form or crystalline form must be free from the other impurities including the unwanted enantiomer, these can be side product and by product of the reaction, degradation products and starting materials. Impurities in final API are undesirable and in extreme cases, might even be harmful to a patient being treated with a dosage form containing the API. Therefore impurities introduced during commercial manufacturing processes must be limited to very small amounts and are preferably substantially absent. These limits are less than about 0.15 percent by weight of each identified impurity and 0.10 % by weight of unidentified and/or uncharacterized impurities. After the manufacture of APIs, the purity of the products, such as (S)- mitiglinide calcium dihydrate is required before commercialization, and in the manufacture of formulated pharmaceuticals. Therefore, pharmaceutical active compounds must be either free from these impurities or contain the impurities in acceptable limits. There is also a need for the isolation, characterization and identification of the impurities and their use as reference markers and reference standard. Thus, the present invention meets the need in the art for a novel, efficient and industrially advantageous process for providing optically pure perhydroisoindole derivatives, particularly (iS)-mitiglinide, which is unique with respect to its simplicity, scalability and involves controlling the steps of the reaction so that predominantly the desired (S)-enantiomer is produced in high yields and purity. The present invention also provides substantially pure (S)-mitiglinide and salts thereof having novel amide impurity in acceptable limit or free from this impurity.

Example 1: Preparation of (R) 4-benzyl-3-(3-phenylpropionv0-oxazolidin-2-one To a solution of (R)-4-benzyloxazolidin-2-one (50 g), 4-dimethylaminopyridine (4.85 g), 3-phenyl propionic acid (55.08 g) in dichloromethane (375 ml) under nitrogen atmosphere at 0-5 ⁰C, dicyclohexylcarbodiimide (975.65 g) was added. The temperature was slowly raised to 25-30 ⁰C and stirring was continued until no starting material was left as was confirmed by thin layer chromatography. Dicyclohexylurea formed during the reaction was filtered, washed with dichloromethane (200 ml) and the filtrate was washed with saturated solution of sodium bicarbonate (500 ml). The solution was dried over sodium sulphate and solvent was distilled off to obtained crude product which was purified from methanol (200 ml) at 10-15 °C and washed with methanol (50 ml) to obtain 81.0 g of the title compound. Example 2: Preparation of 3(5)-benzyl-4-(4-(J?)-benzyl-2-oxo-oxazolidin-3-yl)-4-oxo-butyrϊc acid tert-butyl ester

To a solution of (/?)-4-benzyl-3-(3-phenyl-propionyl)-oxazolidin-2-one (150 g) in anhydrous tetrahydrofuran (1.5 It) was added a solution of sodium hexamethyldisilazane (462 ml, 36-38% solution in tetrahydrofuran) with stirring at -85 to -95 ⁰C for 60 minutes. Tert-butyl bromo acetate (137.5 g) in tetrahydrofuran (300 ml) was added to reaction mass and then stirred to 60 minutes at -85 to -95 ⁰C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ammonium chloride solution (10%, 2.0 It) and extracted with ethyl acetate (2×750 ml). The combined organic layer was washed with demineralized water (1×750 ml) and dried over sodium sulphate. The solvent was evaporated under reduced pressure to obtain oily residue which was stirred with mixture of n-hexane (100 ml) and isopropyl alcohol (100 ml) at Oto -5⁰C, filtered and dried under vacuum to obtain 153.12 g of title compound having chemical purity 99.41%, chiral purity 99.91% by HPLC, [α]_D ²⁰: (-)97.52° (c = 1, CHCl₃) and M.P. : 117.1-118.2⁰C.

Example 3: Preparation of 3(5)-benzyl-4-(4(i?)-benzyl-2-oxo-oxazolidin-3-yl)-4-oxobutyric acid Trifluoroacetic acid (100 g) was added to a solution of 3(5)-benzyl-4-(4-(/?)-benzyl-2-oxo-oxazolidin-3- yl)-4-oxobutyric acid tert-butyl ester (100 g) in dichloromethane (700 ml) at 25 ⁰C and mixture was stirred further for about 12 hours ( when TLC indicated reaction to be complete). The reaction mixture was poured in to ammonium chloride solution (10%, 500 ml). The dichloromethane layer was separated and aqueous layer was extracted with dichloromethane (2 x 250 ml). The combined organic layer was dried over sodium sulphate and evaporated under reduced pressure to obtain title compound. The crude product was recrystallized from a mixture of ethyl acetate: n-hexane (1:4, 500 ml) to obtain 78.75g of the title compound having purity 99.56% by HPLC and M.P.: 145.9-146.4⁰C.

Example 4: Preparation of (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-vI)-4-(hexahydro- isoindolin-2-yl)-butane-l,4-dione

To a solution of 3(5)-benzyl-4-(4-(/?)-benzyl-2-oxo-oxazolidin-3-yl)-4-oxo-butyric acid (50 g) in anhydrous dichloromethane (1.25 It) was added triethylamine (50 ml) with stirring at -20 to -30 ⁰C and the stirred for 15 minutes. A solution of isobutylchloroformate (37.50g) in anhydrous dichloromethane (50 ml) was added at -20 to -30 ⁰C and stirred for 60 minutes. Thereafter, a solution of cis- hexahydroisoindoline (32.50 g) in anhydrous dichloromethane (50 ml) was slowly added by maintaining temperature -20 to -30⁰C. After the completion of the reaction (monitored by HPLC), the mixture was successively washed with 0.5N hydrochloric acid solution (500 ml), brine (300 ml) and dried over sodium sulphate. The solvent was evaporated under reduced pressure to obtain 102.0 g of the title compound having purity 94.39% by HPLC.

Example 5: Purification of r2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro- isoindolin-2-vD-butane-l,4-dione

To the crude (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)- butane- 1,4-dione (51.0 g) was added methanol (150 ml) and the mixture was stirred for 5 hours at 0 to 5 ⁰C. Solid that precipitated out was filtered, slurry washed with cold methanol (25 ml) and dried at 45 -50 ⁰C under vacuum to obtain 28.80 g of pure title compound as a crystalline solid having purity of 99.71% by HPLC and M. P.: 104.1-105.7⁰C.

Example 6: Preparation of calcium salt of (-SVmitiglinide. Step-1: Preparation of (-SVmitiglinide

(2S)-2-Benzyl- 1 -((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)-butane- 1 ,4-dione (28.0 g) was dissolved in tetrahydrofuran (196 ml) and a mixture of lithium hydroxide monohydrate (3.51 g) in demineralized water (56 ml) and hydrogen peroxide (40% solution, 5.5 ml) was added with stirring at 0 to 5 ⁰C over a period of 30 minutes. The reaction mixture was further stirred at 0 to 5 ⁰C till the completion of the reaction. After the completion of the reaction (monitored by TLC), the reaction was quenched with the addition of cooled sodium meta-bisulphate solution (25%, 168 ml) at 0 to 10 ⁰C. The reaction mixture was extracted with ethyl acetate (2×112 ml), the layers were separated and the aqueous layer was discarded. The HPLC analysis of the aqueous layer shows 0.77% of amide impurity. The ethyl acetate layer was then extracted with aqueous ammonia solution (4%, 2×40 ml). The layers were separated and the aqueous layer was further extracted with ethyl acetate (2×280 ml). Combined ethyl acetate layer was discarded. This aqueous layer (280 ml) was used as such in the next stage. The aqueous layer display purity 96.19 % by HPLC and amide impurity 0.04% by HPLC. Step-2: Preparation of calcium salt of dSVmitiglinide

To the above stirred solution of (S)-mitiglinide in water and ammonia(280 ml), methanol (168 ml) was added, followed by calcium chloride (4.48 g) dissolved in demineralized water (56 ml) at ambient temperature and the mixture was stirred for 2 hours. The resulting precipitate was filtered, successively slurry washed with water (3 x 140 ml) and acetone (2 x 70 ml) and dried at 45⁰C -50⁰C under vacuum to obtain 16.1 g of title compound having purity 99.67% by HPLC and amide impurity 0.01% by HPLC. The title product was re-precipitated from a mixture of methanol and water and dried to obtain pure title compound.

Example 7: Preparation of (.SVmitiglinide

To a solution of (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)- butane- 1,4-dione (50 g) in tetrahydrofuran (350 ml) was added a solution of lithium hydroxide monohydrate (8.65 g) in demineralized water (100 ml) and hydrogen peroxide (30% w/w, 40 ml) with stirring at 5 to 10 ⁰C over a period of 15 minutes. After the completion of reaction, sodium meta- bisulphate solution (40%, 500 ml) was added to the reaction mixture and the mixture was extracted with ethyl acetate (2 x 250 ml). The organic layer was dried over sodium sulphate and evaporated under vacuum to obtain 45.5 g of title compound having 35 % of R-benzyl oxozolidin-2-one as impurity. Example 8: Purification of (.S)-mitiglinide

Aqueous ammonia solution (4%, 300 ml) was added to the crude (5)-mitiglinide (30 g) and stirred. The reaction mixture was washed with ethyl acetate (3 x 300 ml). Thereafter the reaction mixture was acidified to pH 1 to 2 with IN hydrochloric acid solution (250 ml) and extracted with ethyl acetate (2 x 150 ml). The layers were separated and ethyl acetate layer was washed with demineralized water (2 x 150 ml), dried over sodium sulphate and then evaporated under reduced pressure to obtain 16.2 g of pure (5)-mitiglinide having purity 95.55% by HPLC Example 9: Preparation of calcium salt of (S)-mitiglinide

To a solution of (<S)-mitiglinide (15 g) in water (150 ml) and aqueous ammonia solution (25%, 15 ml) at 25 to 30 ⁰C, a solution of calcium chloride (7.5 g) in demineralized water (37.5 ml) was added. The mixture was stirred for 1 hour to precipitate the calcium salt of (5)-mitiglinide dihydrate. The resulting precipitate was filtered, slurry washed with water (3 x 150ml) and dried at 45 to 50 ⁰C to obtain 13.25 g of the title compound having purity of 98.84% by HPLC. Example 10: Purification of calcium salt of (5)-mitiglinide

(iS)-mitiglinide calcium (10 g) was dissolved in dimethylformamide (100 ml). This is followed by the addition of demineralized water (500 ml) at 25 to 30 ⁰C. The mixture was stirred for 30 minutes. The precipitated solid was filtered, washed with water (10x 50ml) and dried at 45 to 50 ⁰C under vacuum to obtain 8g of pure title compound as a crystalline solid having purity of 99.62% by HPLC. Example 11: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium (2.0 g) was dissolved in tetrahydrofuran (20 ml) and filtered to remove undissolved and suspended particles. The solvent was then evaporated under vacuum to obtain a powder which was then dried under vacuum at 40-60⁰C to obtain 1.70 g of the title compound. Example 12: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium (2.0 g) was dissolved in dichloromethane (30 ml) and filtered to remove undissolved and suspended particles. The solvent was then evaporated under vacuum to obtain a powder which was then dried under vacuum at 40-60⁰C to obtain 1.64 g of the title compound. Example 13: Preparation of amorphous mitiglinide calcium

Mitiglinide (2.0 g) was dissolved in methanol (20 ml) and methanolic ammonia (5.0 ml) solution was added to it. The solution was stirred at 25-30 ⁰C and calcium chloride (1.5 g) dissolved in methanol was mixed with the solution of mitiglinide and ammonia in methanol and the solution was filtered to remove the suspended particles. The solvent was then evaporated under vacuum to obtain a powder which was then dried under vacuum at 40-60⁰C to obtain 1.9 g of the title compound. Example 14: Preparation of amorphous mitiglinide calcium

Mitiglinide (2.0 g) was dissolved in dichloromethane (20 ml) and aqueous ammonia (3.6 ml, 25 % solution) was added to it. The solution was stirred at 25-30⁰C and solid calcium chloride (1.5 g) was mixed with the solution of mitiglinide and ammonia in dichloromethane and the solution warmed at 30 – 35 ⁰C. The solution was washed with water (2 xlO ml) and the clear solution was dried over sodium sulfate, filtered and evaporated under vacuum and finally dried at under vacuum at 40-60 ⁰C to obtain 1.75 g of the title compound.

Example 15: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium dihydrate (2.0 g) was dissolved in ethyl acetate (30 ml) and filtered to remove undissolved and suspended particles. Approimately. 60 % of the solvent was distilled off under vacuum to obtain a stirrable solution. The solution was then cooled to 15-2O⁰C, mixed with n-heptane (20 ml) and the mixture was stirred for 30 minutes. The resulting solid was filtered, washed with n-heptane and dried under vacuum at 45-60⁰C to yield 1.72 g of the title compound. Example 16: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium (2.Og) was dissolved in dichloromethane (30 ml) and filtered to remove undissolved and suspended particles. Approximately 60 % of the solvent was distilled off under vacuum to obtain a stirrable solution. The solution was then cooled to 15-20⁰C and mixed with diisopropyl ether (20 ml). The mixture was stirred for 30 minutes and the resulting solid was filtered, washed with diisopropyl ether and dried under vacuum at 45-60⁰C to obtain 1.70 g of the title compound. Example 17: Preparation of amorphous mitiglinide calcium

Mitiglinide (2.0 g) was dissolved in dichloromethane (20 ml) and aqueous ammonia (3.6 ml, 25 % solution) solution was added to it. The solution was stirred at 25-30 ⁰C and mixed with solid calcium chloride (1.5 g) and the solution warmed at 30-35 ⁰C and stirred for 30 minutes. The solution was washed with water (2 x 10 ml) and the clear solution was dried over sodium sulfate, and filtered. Approximately 60% of the solvent was distilled off under vacuum and the resulting viscous oil was cooled to 10-15 ⁰C and mixed with diisopropyl ether (50 ml). The reaction mixture was stirred for 30-35 minutes and the resulting solid was filtered and dried at 40-60⁰C to obtain 1.75 g of the title compound. Example 18: Conversion of amorphous mitiglinide calcium into crystalline mitiglinide calcium A suspension of amorphous mitiglinide calcium in diisopropyl ether (30 ml) was stirred for 2 hours at 25- 30⁰C, filtered and dried under vacuum at 45-60⁰C to obtain crystalline form of mitiglinide calcium. Example 19: Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (2.5 g) in water (2.5 ml), aqueous ammonia solution (approx 25%, 4.0 ml) and acetonitrile (2.5 ml) at 10-15⁰C, calcium chloride (1.32 g) dissolved in demineralized water (15 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 25 ml) and acetone (2 x 5 ml) and dried at 45-50⁰C under vacuum to obtain 2.12 g of title compound having purity: 99.72 % by HPLC.

Example 20: Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (2.5 g) in water (2.5 ml), aqueous ammonia solution (approx 25%, 4.0 ml) and tetrahydrofuran (2.5 ml) at 10-15⁰C, calcium chloride (1.32 g) dissolved in demineralized water (15 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 25 ml) and acetone (2 x 5 ml) and dried at 45-50⁰C under vacuum to obtain 1.95 g of title compound having purity: 99.52 % by HPLC.

Example 21; Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (30.0 g) in water (300 ml), aqueous ammonia solution (approx 25%, 48 ml) and acetone (300 ml) at 10-15⁰C, calcium chloride (15.8 g) dissolved in demineralized water (180 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 300 ml) and acetone (2 x 60 ml) and dried at 45-50⁰C under vacuum to obtain 24.32 g of title compound having purity: 99.42 % by HPLC.

Example 22: Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (3.0 g) in water (30 ml), aqueous ammonia solution (approx 25%, 4.8 ml) and isopropyl alcohol (300 ml) at 10-15⁰C, calcium chloride (1.58 g) dissolved in demineralized water

(18 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 30 ml) and acetone (2 x 6 ml) and dried at 45-50⁰C under vacuum to obtain 1.92 g of title compound having purity: 99.65 % by HPLC.

Example 23: Preparation of (2S)-2-benzyWV-((lR)-l-benzyl-2-hydroxy-ethyl)-4-(hexahvdro- isoindolin-2-yl)-4-oxo-buryramide

To a solution of (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)- butane-l,4-dione (20.0 g) in tetrahydrofuran (140 ml), a solution of lithium hydroxide monohydrate

(3.43 g,) in demineralized water (40 ml) was added and the reaction mixture was refluxed for 4 hours till the completion of the reactions (monitored by thin layer chromatography). After the completion of the reaction, the reaction mixture was poured into demineralized water (100 ml) and extracted with ethyl acetate (2 x 80 ml). The combined organic layer was washed with water (80 ml) and dried over sodium sulphate. The solvent was evaporated under reduced pressure to give residue which was stirred in isopropyl alcohol at 0-5 ⁰C for 5 hours. The mixture was filtered and then dried at 40-45 ⁰C under vacuum to obtain 12.48 g of title compound having purity 99.77 % by HPLC. Melting point = 77 – 80⁰C.

PAPER

An Effective and Convenient Method for the Preparation of KAD-1229

Helvetica Chimica ActaVolume 87, Issue 8, Version of Record online: 27 AUG 2004

PAPER

asian journal of chemistry asian journal of chemistry

PATENT

https://www.google.com/patents/CN104311471A?cl=en

Mitiglinide calcium (mitiglinide calcium), the chemical name (2S) -2_ benzyl-3- (cis – hexahydro-2-isoindoline-carbonyl) propionic acid calcium salt dihydrate , for the treatment of type II diabetes. Kissei by Japanese pharmaceutical company research and development, and for the first time on sale in Japan in May 2004. Mitiglinide calcium is the second repaglinide, nateglinide after the first three columns MAG urea drugs, are ATP-dependent potassium channel blocker, is a derivative of phenylalanine, and its mechanism Similar sulfonylureas, but a faster onset of action and short half-life, is conducive to reducing postprandial blood glucose in diabetic patients, and avoid continuous glucose-induced low blood sugar, with the “in vitro pancreas” reputation.

郑德强 etc. on “Food and Drug” magazine was first disclosed the synthesis of calcium Mitiglinide, this method dimethyl succinate and benzaldehyde for raw materials, Stobble condensation, hydrolysis, dehydration anhydride, cis – perhydro isoindole reduced to give racemic acid after condensation, and then split, and salt get Mitiglinide calcium. Specific synthetic route the following equation. The method is relatively complex, in the preparation process to generate half of the unwanted enantiomer, which will waste a lot of cis – perhydro isoindole, and in the preparation of cis – to use science as a whole hydride hydrogen isoindole time reducing agent, the operation is more complicated, the cost is relatively high, and the chiral amine as a resolving agent split, the yield is low.

 The patent discloses a CN201010573666 diethyl succinate and benzaldehyde, condensation occurs Stobble sodium ethoxide in ethanol and then hydrolyzed benzylidene succinic acid, succinic acid benzylidene get by catalytic hydrogenation DL-2-benzyl succinic acid, DL-2-benzyl succinic acid by (R) – a chiral amine resolving to give (S) -2- benzyl succinic acid, (S) -2- benzyl succinic acid anhydride to generate its role in the acetic anhydride, and the resulting acid anhydride and cis – hexahydro isoindole reaction of Mitiglinide acid, calcium chloride and ammonia most 后米格列奈 acid reacts with calcium Mitiglinide dihydrate. The synthesis route following formula. This method effectively avoids the expensive intermediate cis – perhydro isoindole waste, reduce costs, but still amounted to a six-step synthesis route much so that the reagent type, long cycle, low yield, and direct use in the synthesis process Sodium block protonated reagent preparation sodium methylate, generate a lot of flammable hydrogen gas, limiting the industrial application of the method.

The present invention solves is to overcome the existing routes that exist in step lengthy reagent variety, low yield, long cycle, high cost, not suitable for industrial production shortcomings. The present invention provides the following formula preparation process route mitiglinide calcium, organic solvent for this preparation method uses less synthesis process is simple, high yield, good purity, suitable for industrial production.

An improved Mitiglinide calcium industrialized preparation method comprises the following steps: Step 1: Preparation of 2-benzylidene succinic acid; 2 steps: (S) prepared _2_ section succinic acid; Step 3: 2- (S) – section group _4_ oxo – (cis – perhydro isoindol-2-yl) butyric acid; Step 4: Preparation Mitiglinide calcium. Characterized in that: in step 1, using commercially available reagents protonated organic bases, protonation process using an organic alkali solution was slowly feeding methods. Step 2 chiral asymmetric reduction. Step 3 fails anhydride using direct selective amidation. Step 4 beating impurities using an aqueous solvent, prepared mitiglinide calcium dihydrate purification method.

The preparation step 1, using a commercially available organic bases as sodium methoxide or sodium ethoxide protonation agent. As optimization program, feeding method using sodium methoxide or sodium ethoxide solution formulated as the corresponding alcohol and the corresponding dialkyl succinate protonating a nucleophilic substitution reaction.

 The preparation method described in Step 2, the use of Ru with BINAP homogeneous catalyst Ru (OAc) 2 [(S) -BINAP] as a chiral asymmetric synthesis of chiral reducing reagent.

The steps of the preparation method 3, using ethyl acetate as a reaction solvent, acid binding agent triethylamine do, imidazole and thionyl chloride selective amidation reagent, for cis – perhydro isoindole conduct Selective condensation title intermediate.

 The step of preparing said 4, mitiglinide calcium crude product was slurried in 95% ethanol by suction, after simple preparation of high purity mitiglinide calcium dihydrate.

 More specifically, the industrialized Mitiglinide calcium preparation, the following steps: Step 1: Preparation of succinate 2_ Benzylidene

Sodium methoxide (sodium ethoxide) was dissolved in methanol (ethanol), was added dropwise to dimethyl succinate (ethyl) ester, was heated at reflux for 30min, benzaldehyde was added dropwise under reflux, stirring at reflux completed the dropwise 3~5h, drops adding an aqueous solution of 4N NaOH dropwise Bi refluxed 4~6h, cooled to room temperature, adjusted with 6N HCl San PH 2, a solid precipitated, centrifuged, and dried to give the title intermediate 1. Step 2: Preparation of (S) -2- acid, benzyl butyl

Intermediate 1, methanol, and Ru (OAc) 2 [(S) -BINAP] into the reactor, the reactor with N2 the replacement air after heating to 50 ° C, a hydrogen pressure through 10h, cooled, filtered, The filtrate was concentrated to dryness to give the title intermediate 2. Step 3: 2- (S) – benzyl-4-oxo – (cis – perhydro isoindol-2-yl) butyric acid

Ethyl acetate was added to the reactor, triethylamine, imidazole and Intermediate 2, was stirred and cooled to -15~-5 ° C, was added dropwise thionyl chloride addition was complete, the -15 ° C~_5 ° C Under continued stirring 6h, a solution of cis – perhydro isoindole, drip completed, stirred at room temperature overnight, the reaction mixture was added IN hydrochloric acid, stirred Ih, separation, and the organic layer was washed with sodium hydroxide solution to extract IN The combined aqueous layer was washed with a small amount of ethyl acetate, the aqueous layer was adjusted with IN hydrochloric acid and the PH = 3, the aqueous layer was extracted with ethyl acetate, the organic layers combined, washed with water and saturated brine, and the organic layer was dried over anhydrous Na2SO4, filtered and the filtrate concentrated under reduced pressure to obtain the objective compound 3 billion Step 4: Preparation of calcium Mitiglinide

The 3 was dissolved in ethanol, was added 2N sodium hydroxide solution, after mixing the solution was added dropwise a 10% aqueous solution of calcium chloride, the reaction mixture was stirred vigorously 3~5h, ice-cooled, filtered, the filter cake with 95% ethanol beating crystallization, filtration, and dried in vacuo to give the title compound I.

Accordingly, the present invention is a method for preparing mitiglinide calcium has the following advantages:

1, Step 1, using commercially available sodium methylate (sodium ethanol) instead of sodium block as a proton agent, effectively avoid the risk of sodium block formed during the reaction a lot of flammable hydrogen gas, industrial production safer. Another use dropping protonated reagent feeding method can effectively avoid succinic acid alkyl ester of two methylene groups are protonated and reduce the incidence of side effects, so that the yield increased by nearly 20%.

 2, Step 2, the selective reduction of chiral reagent (S) -BINAP instead of the original route after the first split reduction method, not only simplifies the reaction step, but low yield while avoiding split It leads to the risk of an increase in cost.

3, Step 3, the fixed selective amidation reaction conditions instead of the original first into anhydride after amidation reaction that simplifies the reaction steps to reduce the unit operations, shortening the production cycle, improve production efficiency.

4, Step 4, by using an aqueous solution of calcium Mitiglinide ethanol refining crude beating, then dried under reduced pressure to control the moisture content and reduce the difficulty of the operation, more conducive to industrial production.DETAILED DESCRIPTION The following examples further illustrate the invention, but the present invention is not limited thereto. Example One Step I: Preparation 2_ benzylidene succinic acid Sodium methoxide (9kg) and methanol (48L) into the 100L reactor, stirring to dissolve, into the high slot 50L. The dimethyl succinate (20kg) into the 200L reaction vessel, heated to reflux, methanol was added dropwise a solution of fast high tank of sodium methoxide, refluxed for reaction completion dropwise 30min, was added dropwise under reflux benzaldehyde (10. 9kg) dropwise with stirring at reflux completed 3~5h, HPLC detection benzaldehyde completion of the reaction, a solution of aqueous 4N NaOH (38L), Bi dropwise refluxed 4~6h, cooled to room temperature, 2, adjusted with 6N HCl and the precipitated solid was San PH, centrifugation, and dried in vacuo to give a pale yellow solid 19kg, i.e. an intermediate, yield 90%. Step 2: Preparation of (S) -2- butyric acid benzyl 200L detecting a high pressure hydrogenation reactor airtight, Intermediate I (19kg), methanol (95L) containing 5% Ru (0Ac) 2 [(S ) -BINAP] molecular sieve (SBA-15) supported catalyst (0. 95kg, homemade) into the reactor, purge the inside of the reactor with N2 atmosphere, followed by heating to 50 ° C, atmospheric pressure hydrogen-10h, cooled, filtered and the filtrate was concentrated to dryness under reduced pressure, the resulting solid was recrystallized from ethyl acetate and dried in vacuo to give an off-white solid 15. 5kg, i.e. intermediate 2, yield 81%, chiral purity 90. 5% θ. θ .. Step 3: 2- (S) – benzyl-4-oxo – (cis – perhydro isoindol-2-yl) butyric acid in 500L reaction vessel was charged with ethyl acetate (225L), triethylamine (1.8kg), imidazole (9. 8kg) and Intermediate 2 (15kg), stirred and cooled to -KTC, was added dropwise thionyl chloride (17. 2kg), the addition was complete, the -KTC~-5 ° C under Stirring was continued for 6h, a solution of cis – perhydro isoindole (9kg), drip completed, the reaction was stirred at room temperature for 18h, the reaction mixture was added IN HCl (150L) was stirred Ih, liquid separation, the organic layer was washed with IN sodium hydroxide solution (100LX3) extracted aqueous layers were combined, washed with ethyl acetate (50L) with, water layer was washed with IN of hydrochloric acid adjusted to PH = 3, the aqueous layer was extracted with ethyl acetate (IOOmLX 3), the combined organic layers , saturated brine (50LX 3) was washed, and the organic layer was dried over anhydrous Na2SO4, filtered, and the filtrate was concentrated under reduced pressure to give an oil 19. 8kg, i.e. Intermediate 3 Yield: 87%. Step 4: Preparation of mitiglinide calcium Intermediate 3 (. 19 8kg) and absolute ethanol (99L) into the 200L reactor, and stirred to dissolve, was added 2N sodium hydroxide solution (35L), minutes after mixing Batch into the high slot. The 500L reaction vessel was added 5% aqueous calcium chloride solution (155L), stirring was added dropwise a solution of the high slot, dropwise with vigorous stirring the reaction completion 3~5h, centrifuged, the cake was washed with 95% ethanol (99L) was recrystallized beating, centrifugation and dried in vacuo (50 ° C / 0. 09MPa), to give the title compound I 16. lkg, yield 73%.

PATENT

https://www.google.com/patents/CN102424664A?cl=en

Mitiglinide calcium Phenylalanine belong chiral compound synthesis routes according to different methods of constructing chiral center has the following three synthetic process:

① split method Document: CN 102101838A, CN 1844096, etc.)

In this method, diethyl succinate and benzaldehyde by Mobbe condensation, hydrolysis, dehydration anhydride, and after cis-hydrogenated isoindole condensation is reduced to give racemic acid, and then split, and salt to give Mitiglinide calcium. The first method step condensation reaction impurities, product separation and purification difficult, finally resolving the yield is low. This method is also a lack atom economy.

 ② asymmetric hydrogenation Document tetrahedron Letters, 1987,28 (17), 1905-1908; Tetrahedron Letters, 1989,30 (6), 735-738)

[0027] This method requires expensive rhodium complexes (Rh, (2S, 4Q-N_-butoxycarbonyl-4-diphenylphosphino _2_ diphenylphosphino-2-diphenylphosphino methylpyrrolidine alkyl), making the production cost is greatly improved, and the need for high-pressure hydrogenation reaction, is not conducive to industrial production.

③ chiral method Document: CN 1680321A)

The method uses phenylalanine as chiral starting materials, after diazotization, nucleophilic substitution, high temperature decarboxylation and condensation reaction product. Wherein the decarboxylation temperature is too low yield, making the overall process costs.

DISCLOSURE

The object of the present invention is to provide a simple, effective and easy-to-operate preparation Mitiglinide calcium.

The present invention provides a process for the preparation of calcium Mitiglinide, the synthesis route is as follows:

 Step 1: D- phenylalanine in the acid hydrolysis of formula (¾ 2- hydroxy acid;

Step 2: formula (¾ 2- hydroxy acid under basic conditions to give protected hydroxyl sulfonate of formula (¾-hydroxyphenyl propionic acid ester;

 Step 3: The formula (¾-hydroxyphenyl propionic acid ester in the acid-catalyzed carboxyl ester-protected formula (4) phenylalanine methyl sulfonate carboxylate;

 Step 4: cis-hydrogen isoindole synthesis formula (6) perhydro isoindole halide;

Step 5: Under alkaline conditions, the formula ⑷ formula (6) nucleophilic substitution reaction formula (5) Mitiglinide acid

Step 6: Under alkaline conditions, the formula (¾ Mitiglinide ester hydrolysis to the calcium salt of formula (1) Mitiglinide calcium.

 Preferably, the specific steps include:

 Step 1: (D) – phenylalanine hydrolysis in a strong acid of formula (2) 2-hydroxyphenyl propionic acid

 In (D) – phenylalanine as a starting material, in the presence of a strong acid such as sulfuric acid, _5 ° C _5 ° C hydrolysis, to give Formula (2) 2-hydroxyphenyl propionic acid White solid.

 Step 2: The formula (¾ 2- hydroxy acid under basic conditions to protect the hydroxyl group sulfonic acid ester of formula (¾-hydroxyphenyl propionic acid ester

2-hydroxyphenyl propionic acid in an organic base such as triethylamine or pyridine, or an inorganic base such as sodium bicarbonate, sodium carbonate or potassium carbonate effect, p-hydroxybenzoic acid ester protecting performed, the protecting group used is an aliphatic or aromatic sulfonic acid group such as mesylate, tosylate or p-toluenesulfonic acid group, a sulfonic acid group is preferably methyl group or p-toluenesulfonic acid.

Step 3: Protect formula formula (¾-hydroxyphenyl propionic acid ester in the acid-catalyzed carboxyl ester group (4) benzenepropanoic

MitigIinide1 (I) carboxylic acid ester sulfonate

In the catalytic acid carboxyl benzenepropanoic acid ester group protection, the use of alcohol may be fatty alcohols or aromatic alcohols, preferably ethanol, t-butanol or benzyl alcohol.

 Step 4: cis-hydrogen isoindole synthesis formula (6) perhydro isoindole halide

 In the synthesis of perhydro isoindole halide in the haloacetyl halide can be used chloroacetyl chloride, bromoacetyl chloride or bromoacetyl bromide, chloroacetyl chloride is preferred.

 Step 5: Under alkaline conditions, (4) and (6) a nucleophilic substitution reaction formula (¾ Mitiglinide acid

 Under the conditions of a strong base, such as sodium alkoxide such as sodium ethoxide or sodium methylate, perhydro isoindole halide and phenylalanine sulfonate nucleophilic substitution reaction Mitiglinide ethyl reaction temperature of -10 ° C -25 ° c, preferably 0 ° C.

Step 6: Under alkaline conditions, the formula (¾ Mitiglinide ester hydrolysis to the calcium salt of formula (1) calcium Mitiglinide

Ethyl mitiglinide under basic conditions such as sodium hydroxide, potassium hydroxide, or an amine (ammonia) in the presence of an aqueous solution of calcium chloride, and hydrolyzed as calcium salt, in aqueous solution under conditions of heavy alcohol crystallization, high purity mitiglinide calcium.

 The present invention and the prior art comparison, has the following advantages:

1, to find an innovative high-yield process for preparing calcium Mitiglinide route, a total yield of 47%;

2, with respect to the routing methods reported in the literature, the optical yield doubled, ee greater than 99%;

3. The process route of the raw materials are cheap, readily available, avoiding costly chiral resolving agents or the use of a catalyst;

 4. The process route mild conditions, high temperature decarboxylation overcome the harsh reaction conditions.

 In the present invention, (D) – phenylalanine as a starting material, after diazotization, a hydroxyl group and a carboxyl group protected, nucleophilic substitution, hydrolysis and other reactions prepared mitiglinide calcium, high yield. The present invention provides a process used by a wide range of raw materials, low prices, the total yield of 47%, optical purity greater than 99%, and mild reaction conditions, the reaction process is simple, avoid the literature, such as split, high-pressure hydrogenation method low yield, long reaction steps and other shortcomings, but also to overcome the harsh conditions of high temperature reaction deacidification, etc. for preparation and production of calcium Mitiglinide provides a new choice.

The process route mild conditions, high temperature decarboxylation overcome the harsh reaction conditions.

 In the present invention, (D) – phenylalanine as a starting material, after diazotization, a hydroxyl group and a carboxyl group protected, nucleophilic substitution, hydrolysis and other reactions prepared mitiglinide calcium, high yield. The present invention provides a process used by a wide range of raw materials, low prices, the total yield of 47%, optical purity greater than 99%, and mild reaction conditions, the reaction process is simple, avoid the literature, such as split, high-pressure hydrogenation method low yield, long reaction steps and other shortcomings for Mitiglinide calcium preparation and production of a new choice.

Preferably, in the above embodiment, each step may be the following alternative, the embodiment can achieve the same advantageous effects to a third embodiment of embodiment:

 Step 1: (D) – phenylalanine in the acid hydrolysis of formula (¾ 2- hydroxy acid

 In (D) – phenylalanine as a starting material, in the presence of sulfuric acid, -50C _5 ° C hydrolysis, to give Formula O) 2-hydroxyphenyl propionic acid White solid.

Step 2: formula (¾ 2- hydroxy acid under basic conditions to give protected hydroxyl sulfonate of formula C3) hydroxyphenyl propionic acid ester

2-hydroxyphenyl propionic acid in an organic base such as triethylamine or pyridine, or an inorganic base such as sodium bicarbonate, sodium carbonate or potassium carbonate effect, p-hydroxybenzoic acid ester protecting performed, the protecting group used is an aliphatic or aromatic sulfonic acid group such as mesylate, tosylate or p-toluenesulfonic acid group, a sulfonic acid group is preferably methyl group or p-toluenesulfonic acid.

Step 3: Formula C3) hydroxyphenyl propionic acid ester in the acid-catalyzed carboxyl ester-protected formula (4) phenylalanine methyl sulfonate carboxylate [0118] In the acid-catalyzed, styrene-acrylic acid ester-protected carboxy, the use of alcohol may be fatty alcohols or aromatic alcohols, preferably ethanol, t-butanol or benzyl alcohol.

 Step 4: cis-hydrogen isoindole synthesis formula (6) perhydro isoindole halide

In the synthesis of perhydro isoindole halide in the haloacetyl halide can be used chloroacetyl chloride, bromoacetyl chloride or bromoacetyl bromide, chloroacetyl chloride is preferred.

Step 5: Under alkaline conditions, the formula ⑷ formula (6) nucleophilic substitution reaction formula (5) Mitiglinide acid

Under the conditions of a strong base, such as sodium alkoxide such as sodium ethoxide or sodium methylate, perhydro isoindole halide and phenylalanine sulfonate nucleophilic substitution reaction Mitiglinide ethyl reaction temperature of -10 ° C -25 ° c, preferably 0 ° C.

 Step 6: Under alkaline conditions, the formula (¾ Mitiglinide ester hydrolysis to the calcium salt of formula (1) calcium Mitiglinide

 Ethyl mitiglinide under basic conditions such as sodium hydroxide, potassium hydroxide, or an amine (ammonia) in the presence of an aqueous solution of calcium chloride, and hydrolyzed as calcium salt, in aqueous solution under conditions of heavy alcohol crystallization, high purity mitiglinide calcium.

Patent

https://www.google.com/patents/CN103724253A?cl=en

bis [(2s) -2- benzyl-3- (cis – hexahydro isoindole-2-carbonyl) propionic acid] monocalcium dihydrate (mitiglinide calcium), the formula C38H48CaN206.2Η20 English called Mitiglinide Calcium Hydrate, structural formula (I) as

 Mitiglinide Calcium is synthesized by Japan Orange Health Pharmaceutical Co., Ltd., in April 2004 in Japan, for through diet and exercise therapy can effectively control high blood sugar in type II diabetes patients.Mitiglinide calcium is the second repaglinide, nateglinide third after the United States and Glenn urea drugs belong phenylalanine derivatives. By closing ΑΤΡ Mitiglinide calcium-dependent pancreatic β cell membrane Κ channel, resulting in the Ca flow, increase intracellular Ca concentration of extracellular vesicles containing threshing leaving insulin, thereby stimulating the secretion of insulin.And only when the meal will be rapid and transient stimulates the pancreas to secrete insulin, sulphonylureas with the traditional Compared to the rapid onset and short duration of action, inhibition of postprandial hyperglycemia characteristic of type II diabetes, to avoid low blood sugar react, early first- and mild diabetes treatment, and well tolerated.

According to the literature and patent reports, prepared Mitiglinide calcium are the following methods.

 Method I: 2_ (S) _ benzyl succinic acid as raw material, amides, reduction, calcium salt formation Mitiglinide this method, although fewer steps, but the chiral compound materials, expensive , the production cost is high, not suitable for industrial production. References: Sorbera LA, Leeson PA, Castaner RM, et al.Mitiglinidecalcium (KAD-1229) [J] .Drugs Future, 2000,25 (10):. 1034-1042 [0007] Method Two: succinate methyl ester with benzaldehyde for raw materials, Stobble condensation, hydrolysis, dehydration anhydride, cis – perhydro isoindole after condensation is reduced to give racemic acid, and then split into calcium salts and the like have Mitiglinide. This method is relatively complex and condensation reaction impurities, product separation and purification difficult, costly, and chiral separation time yield is low.[Reference: Zheng Dejiang, Liu Wentao, Wu Lihua synthetic calcium Mitiglinide [J] Food and Drug, 2007,9 (11): 13-15]

 Method three: dimethyl succinate and benzaldehyde for raw materials, Stobbe condensation, reduction, split, with p-nitrophenol and dicyclohexyl carbodiimide activated calcium salt formation Mitiglinide This production cost is relatively high, and used column chromatography, suitable for industrial production. References: Synthesis Technology Zhang Hongmei Chen meritorious, Cao Xiaohui Mitiglinide of [J], modern chemicals, 2008,28 (8): 56-59.]

Example 1:

The cis – hexahydro-isoquinoline (250.4g, 2mol), anhydrous potassium carbonate (304.0g, 2.2mol), methylene burn (1000ml) was added to the reaction flask, keeping the temperature 0-5 ° C with vigorous stirring, dropwise acetyl chloride (271.0g, 2.4mol) in dichloromethane (500ml) solution, drip completed, room temperature 2.5h, point board monitoring, reaction complete, additional water 1000ml, organic layer was separated, water (1000ml), saturated brine (1000ml), dried over anhydrous sodium sulfate overnight, dichloromethane was distilled off under reduced pressure to give cis -N- chloroacetyl hexahydro isoindole (2) 357.4g oil close Rate: 88.6%.

The cis -N- chloroacetyl hexahydro isoindole (302.5g, 1.5mol), N_ within phenylpropionyl camphor sulfonamide (573.0g, 1.65mol), 70% sodium hydride (56.6g, 1.65 mol), Ν, Ν- dimethylformamide (900ml) was added to the reaction flask, at 50 ° C, the reaction was stirred vigorously 12h, to give the alkylated product, placed to room temperature before use.

100ml of water was slowly dropped to the above-mentioned system, drip complete, lithium hydroxide (39.5g, 1.65mol), tetrahydrofuran (600ml), at 0-5 ° C under a 30% solution of hydrogen peroxide solution 680ml, drop Albert, was transferred to the reaction was continued at room temperature for 18h, point board monitoring, reaction complete, additional water 1200ml, adjusting the pH to about 2_3, extracted with dichloromethane (900ml X 3), the combined organic phases with saturated brine (1500ml) wash, overnight over anhydrous sodium sulfate, the solvent was distilled off under reduced pressure to give a viscous liquid, to which was added ethyl acetate 250ml, stirred at room temperature, suction filtered, the filter cake with ethyl acetate (150ml) and dried to give (2s) – 2-benzyl-3- (cis – hexahydro isoindole-2-carbonyl) – propionic acid (6) as a white solid 231.8g, two steps yield: 49%. Compound 6 (230g, 0.73mol), water 1150ml, added to the reaction flask. After the whole solution, was added 2mol / L sodium hydroxide solution, 400ml, stirred at rt for 30min, was slowly added dropwise with vigorous stirring chloride (162.0g, 1.46mol) in water (320ml) solution dropwise was completed, the reaction was continued for 1.5h, filtration, water (200ml X 2) washing the filter cake to give a white solid, 60 ° C and dried under reduced pressure to 3h, the filter cake with 95% ethanol (2300ml) recrystallized Mitiglinide calcium (I) 430g, yield: 83.6%, mp: 178 ~ 183 ° C, FAB-MS: m / z316 [M + l] +; [α] D20 = + 5.45 ° (C = 1, methanol) [Document: m.ρ.: 179 ~ 185Ό, [α] d20 = + 5.64 ° (C = L 0, methanol)]; purity: 99.8% [HPLC normalization method : Column C18, mobile phase L OOmol / L potassium dihydrogen phosphate buffered saline – acetonitrile-water (20:35: 30) (adjusted pH = 2.10); detection wavelength 210nm]; iH-NMlUCDCldOOM), δ: 1.1 ~ 1.5 (16Η, m), 1.8 ~ 2.4 (6Η, m), 2.5 ~ 3.1 (14Η, m) 3.3 ~

3.8 (6H, m) 7.4 ~ 7.6 (10H, m); Elemental analysis (%):. C64.68, Η7.35, Ν3.94, Theory: C64.75, Η7.44, Ν3.97 yield : 36.05%, a purity of 99.8%.

PAPER

WEI HUANG，等: “Novel Convenient Synthesis of Mitiglinide“, 《SYNTHETIC COMMUNICATIONS》, vol. 37, no. 13, 3 July 2007 (2007-07-03), pages 2153 – 2157, XP055079498, DOI: doi:10.1080/00397910701392590

http://www.tandfonline.com/doi/abs/10.1080/00397910701392590

Abstract: A novel convenient synthesis of the hypoglycemic agent mitiglinide was developed. (2S)-4-[(3aR,7aS)-Octahydro-2H-isoindol-2-yl]-4-oxo-2-benzyl-butanoic acid (6) was prepared by selective hydrolysis of ethyl 4-[(3aR,7aS)-octahydro-2Hisoindol-2-yl]-4-oxo-2-benzyl-butanoate (5) using a-chymotrypsin; the latter was prepared by a novel facile route from (3aR,7aS)-octahydro-2H-isoindole. The overall yield was 25.6%.

Keywords: a-chymotrypsin, mitiglinide, synthesis

Mitiglinide (calcium bis[(2S)-4-[(3aR,7aS)-octahydro-2H–isoindol-2-yl]-4oxo-2-benzylbutanoate]dihydrate) is a novel oral hypoglycemic agent. It inhibits the adenosine triphosphate (ATP)-sensitive potassium channels in pancreatic b-cells and stimulates insulin release like sulfonylureas,[1] but has a rapid onset and short-lasting hypoglycemic effect as compared with the latter.

Mitiglinide has been synthesized by several related methods that involve optical resolution,[2] asymmetric synthesis,[2a,3] and diasteroselective alkylation using chiral auxiliary.[4]

In a previous article,[2] two optical resolution methods of the key compound racemic acid 4 were reported. One of them involves esterification with optically active alcohols, which are separated into the diastereomers by column chromatogeaphy and hydrolyzed. Only the diastereomeric (S)-Nbenzyl mandelamide ester could be separated; the overall yield was 28%,

The alternative method was optical resolution by optically active bases. The best result was 30.8% yield and 97% ee when using (R)-1-(1-naphthyl)-ethylamine as a base. In this article, we have developed a new optical resolution method of racemic ester 5 by a-chymotrypsin in 45.3% yield; the optical purity of (S)-acid (6) determined by chiral-phase high performance liquid chromatography (HPLC) on Sumichiral

OA3300 was 99.2% ee, and, the method can be used for scale-up preparation.

The synthesis of free acid 6 is shown in Scheme 1. (3aR,7aS)-Octahydro2H-isoindole was chloroacetylated in the presence of Et3N to afford (3aR, 7aS)-2-(chloro-acetyl)-octahydro-2H-isoindole (2), which was condensed with diethyl benzylmalonate followed by hydrolysis and decarbonylation to obtain 4-[(3aR,7aS)-octahydro-2H-isoindol-2-yl]-4-oxo-2-benzyl-butanoic acid (4). The overall yield of the three-step synthesis was 62.9%. The racemic acid (4) was esterified with SOCl2/EtOH to give the corresponding racemic ester (5). The (R)-ester was selectively hydrolyzed by a-chymotrypsin to separate out the (S)-ester, which was subjected to hydrolysis, giving 6.

The overall yield was 28.5% [based on (3aR,7aS)-octahydro-2H-isoindole].

Compound 6 was treated with calcium chloride and 25% ammonium hydroxide to give mitiglinide; after recrystallization from 95% EtOH, the pure product was obtained in 90% yield.

Patent

https://www.google.com/patents/WO2009047797A2?cl=en

EXAMPLES

Example 1: Preparation of (R) 4-benzyl-3-(3-phenylpropionv0-oxazolidin-2-one To a solution of (R)-4-benzyloxazolidin-2-one (50 g), 4-dimethylaminopyridine (4.85 g), 3-phenyl propionic acid (55.08 g) in dichloromethane (375 ml) under nitrogen atmosphere at 0-5 ⁰C, dicyclohexylcarbodiimide (975.65 g) was added. The temperature was slowly raised to 25-30 ⁰C and stirring was continued until no starting material was left as was confirmed by thin layer chromatography. Dicyclohexylurea formed during the reaction was filtered, washed with dichloromethane (200 ml) and the filtrate was washed with saturated solution of sodium bicarbonate (500 ml). The solution was dried over sodium sulphate and solvent was distilled off to obtained crude product which was purified from methanol (200 ml) at 10-15 °C and washed with methanol (50 ml) to obtain 81.0 g of the title compound. Example 2: Preparation of 3(5)-benzyl-4-(4-(J?)-benzyl-2-oxo-oxazolidin-3-yl)-4-oxo-butyrϊc acid tert-butyl ester

To a solution of (/?)-4-benzyl-3-(3-phenyl-propionyl)-oxazolidin-2-one (150 g) in anhydrous tetrahydrofuran (1.5 It) was added a solution of sodium hexamethyldisilazane (462 ml, 36-38% solution in tetrahydrofuran) with stirring at -85 to -95 ⁰C for 60 minutes. Tert-butyl bromo acetate (137.5 g) in tetrahydrofuran (300 ml) was added to reaction mass and then stirred to 60 minutes at -85 to -95 ⁰C. After completion of the reaction (monitored by TLC), the reaction mixture was poured into ammonium chloride solution (10%, 2.0 It) and extracted with ethyl acetate (2×750 ml). The combined organic layer was washed with demineralized water (1×750 ml) and dried over sodium sulphate. The solvent was evaporated under reduced pressure to obtain oily residue which was stirred with mixture of n-hexane (100 ml) and isopropyl alcohol (100 ml) at Oto -5⁰C, filtered and dried under vacuum to obtain 153.12 g of title compound having chemical purity 99.41%, chiral purity 99.91% by HPLC, [α]_D ²⁰: (-)97.52° (c = 1, CHCl₃) and M.P. : 117.1-118.2⁰C.

Example 3: Preparation of 3(5)-benzyl-4-(4(i?)-benzyl-2-oxo-oxazolidin-3-yl)-4-oxobutyric acid Trifluoroacetic acid (100 g) was added to a solution of 3(5)-benzyl-4-(4-(/?)-benzyl-2-oxo-oxazolidin-3- yl)-4-oxobutyric acid tert-butyl ester (100 g) in dichloromethane (700 ml) at 25 ⁰C and mixture was stirred further for about 12 hours ( when TLC indicated reaction to be complete). The reaction mixture was poured in to ammonium chloride solution (10%, 500 ml). The dichloromethane layer was separated and aqueous layer was extracted with dichloromethane (2 x 250 ml). The combined organic layer was dried over sodium sulphate and evaporated under reduced pressure to obtain title compound. The crude product was recrystallized from a mixture of ethyl acetate: n-hexane (1:4, 500 ml) to obtain 78.75g of the title compound having purity 99.56% by HPLC and M.P.: 145.9-146.4⁰C.

Example 4: Preparation of (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-vI)-4-(hexahydro- isoindolin-2-yl)-butane-l,4-dione

To a solution of 3(5)-benzyl-4-(4-(/?)-benzyl-2-oxo-oxazolidin-3-yl)-4-oxo-butyric acid (50 g) in anhydrous dichloromethane (1.25 It) was added triethylamine (50 ml) with stirring at -20 to -30 ⁰C and the stirred for 15 minutes. A solution of isobutylchloroformate (37.50g) in anhydrous dichloromethane (50 ml) was added at -20 to -30 ⁰C and stirred for 60 minutes. Thereafter, a solution of cis- hexahydroisoindoline (32.50 g) in anhydrous dichloromethane (50 ml) was slowly added by maintaining temperature -20 to -30⁰C. After the completion of the reaction (monitored by HPLC), the mixture was successively washed with 0.5N hydrochloric acid solution (500 ml), brine (300 ml) and dried over sodium sulphate. The solvent was evaporated under reduced pressure to obtain 102.0 g of the title compound having purity 94.39% by HPLC.

Example 5: Purification of r2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro- isoindolin-2-vD-butane-l,4-dione

To the crude (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)- butane- 1,4-dione (51.0 g) was added methanol (150 ml) and the mixture was stirred for 5 hours at 0 to 5 ⁰C. Solid that precipitated out was filtered, slurry washed with cold methanol (25 ml) and dried at 45 -50 ⁰C under vacuum to obtain 28.80 g of pure title compound as a crystalline solid having purity of 99.71% by HPLC and M. P.: 104.1-105.7⁰C.

Example 6: Preparation of calcium salt of (-SVmitiglinide. Step-1: Preparation of (-SVmitiglinide

(2S)-2-Benzyl- 1 -((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)-butane- 1 ,4-dione (28.0 g) was dissolved in tetrahydrofuran (196 ml) and a mixture of lithium hydroxide monohydrate (3.51 g) in demineralized water (56 ml) and hydrogen peroxide (40% solution, 5.5 ml) was added with stirring at 0 to 5 ⁰C over a period of 30 minutes. The reaction mixture was further stirred at 0 to 5 ⁰C till the completion of the reaction. After the completion of the reaction (monitored by TLC), the reaction was quenched with the addition of cooled sodium meta-bisulphate solution (25%, 168 ml) at 0 to 10 ⁰C. The reaction mixture was extracted with ethyl acetate (2×112 ml), the layers were separated and the aqueous layer was discarded. The HPLC analysis of the aqueous layer shows 0.77% of amide impurity. The ethyl acetate layer was then extracted with aqueous ammonia solution (4%, 2×40 ml). The layers were separated and the aqueous layer was further extracted with ethyl acetate (2×280 ml). Combined ethyl acetate layer was discarded. This aqueous layer (280 ml) was used as such in the next stage. The aqueous layer display purity 96.19 % by HPLC and amide impurity 0.04% by HPLC. Step-2: Preparation of calcium salt of dSVmitiglinide

To the above stirred solution of (S)-mitiglinide in water and ammonia(280 ml), methanol (168 ml) was added, followed by calcium chloride (4.48 g) dissolved in demineralized water (56 ml) at ambient temperature and the mixture was stirred for 2 hours. The resulting precipitate was filtered, successively slurry washed with water (3 x 140 ml) and acetone (2 x 70 ml) and dried at 45⁰C -50⁰C under vacuum to obtain 16.1 g of title compound having purity 99.67% by HPLC and amide impurity 0.01% by HPLC. The title product was re-precipitated from a mixture of methanol and water and dried to obtain pure title compound.

Example 7: Preparation of (.SVmitiglinide

To a solution of (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)- butane- 1,4-dione (50 g) in tetrahydrofuran (350 ml) was added a solution of lithium hydroxide monohydrate (8.65 g) in demineralized water (100 ml) and hydrogen peroxide (30% w/w, 40 ml) with stirring at 5 to 10 ⁰C over a period of 15 minutes. After the completion of reaction, sodium meta- bisulphate solution (40%, 500 ml) was added to the reaction mixture and the mixture was extracted with ethyl acetate (2 x 250 ml). The organic layer was dried over sodium sulphate and evaporated under vacuum to obtain 45.5 g of title compound having 35 % of R-benzyl oxozolidin-2-one as impurity. Example 8: Purification of (.S)-mitiglinide

Aqueous ammonia solution (4%, 300 ml) was added to the crude (5)-mitiglinide (30 g) and stirred. The reaction mixture was washed with ethyl acetate (3 x 300 ml). Thereafter the reaction mixture was acidified to pH 1 to 2 with IN hydrochloric acid solution (250 ml) and extracted with ethyl acetate (2 x 150 ml). The layers were separated and ethyl acetate layer was washed with demineralized water (2 x 150 ml), dried over sodium sulphate and then evaporated under reduced pressure to obtain 16.2 g of pure (5)-mitiglinide having purity 95.55% by HPLC Example 9: Preparation of calcium salt of (S)-mitiglinide

To a solution of (<S)-mitiglinide (15 g) in water (150 ml) and aqueous ammonia solution (25%, 15 ml) at 25 to 30 ⁰C, a solution of calcium chloride (7.5 g) in demineralized water (37.5 ml) was added. The mixture was stirred for 1 hour to precipitate the calcium salt of (5)-mitiglinide dihydrate. The resulting precipitate was filtered, slurry washed with water (3 x 150ml) and dried at 45 to 50 ⁰C to obtain 13.25 g of the title compound having purity of 98.84% by HPLC. Example 10: Purification of calcium salt of (5)-mitiglinide

(iS)-mitiglinide calcium (10 g) was dissolved in dimethylformamide (100 ml). This is followed by the addition of demineralized water (500 ml) at 25 to 30 ⁰C. The mixture was stirred for 30 minutes. The precipitated solid was filtered, washed with water (10x 50ml) and dried at 45 to 50 ⁰C under vacuum to obtain 8g of pure title compound as a crystalline solid having purity of 99.62% by HPLC. Example 11: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium (2.0 g) was dissolved in tetrahydrofuran (20 ml) and filtered to remove undissolved and suspended particles. The solvent was then evaporated under vacuum to obtain a powder which was then dried under vacuum at 40-60⁰C to obtain 1.70 g of the title compound. Example 12: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium (2.0 g) was dissolved in dichloromethane (30 ml) and filtered to remove undissolved and suspended particles. The solvent was then evaporated under vacuum to obtain a powder which was then dried under vacuum at 40-60⁰C to obtain 1.64 g of the title compound. Example 13: Preparation of amorphous mitiglinide calcium

Mitiglinide (2.0 g) was dissolved in methanol (20 ml) and methanolic ammonia (5.0 ml) solution was added to it. The solution was stirred at 25-30 ⁰C and calcium chloride (1.5 g) dissolved in methanol was mixed with the solution of mitiglinide and ammonia in methanol and the solution was filtered to remove the suspended particles. The solvent was then evaporated under vacuum to obtain a powder which was then dried under vacuum at 40-60⁰C to obtain 1.9 g of the title compound. Example 14: Preparation of amorphous mitiglinide calcium

Mitiglinide (2.0 g) was dissolved in dichloromethane (20 ml) and aqueous ammonia (3.6 ml, 25 % solution) was added to it. The solution was stirred at 25-30⁰C and solid calcium chloride (1.5 g) was mixed with the solution of mitiglinide and ammonia in dichloromethane and the solution warmed at 30 – 35 ⁰C. The solution was washed with water (2 xlO ml) and the clear solution was dried over sodium sulfate, filtered and evaporated under vacuum and finally dried at under vacuum at 40-60 ⁰C to obtain 1.75 g of the title compound.

Example 15: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium dihydrate (2.0 g) was dissolved in ethyl acetate (30 ml) and filtered to remove undissolved and suspended particles. Approimately. 60 % of the solvent was distilled off under vacuum to obtain a stirrable solution. The solution was then cooled to 15-2O⁰C, mixed with n-heptane (20 ml) and the mixture was stirred for 30 minutes. The resulting solid was filtered, washed with n-heptane and dried under vacuum at 45-60⁰C to yield 1.72 g of the title compound. Example 16: Preparation of amorphous mitiglinide calcium

Crystalline mitiglinide calcium (2.Og) was dissolved in dichloromethane (30 ml) and filtered to remove undissolved and suspended particles. Approximately 60 % of the solvent was distilled off under vacuum to obtain a stirrable solution. The solution was then cooled to 15-20⁰C and mixed with diisopropyl ether (20 ml). The mixture was stirred for 30 minutes and the resulting solid was filtered, washed with diisopropyl ether and dried under vacuum at 45-60⁰C to obtain 1.70 g of the title compound. Example 17: Preparation of amorphous mitiglinide calcium

Mitiglinide (2.0 g) was dissolved in dichloromethane (20 ml) and aqueous ammonia (3.6 ml, 25 % solution) solution was added to it. The solution was stirred at 25-30 ⁰C and mixed with solid calcium chloride (1.5 g) and the solution warmed at 30-35 ⁰C and stirred for 30 minutes. The solution was washed with water (2 x 10 ml) and the clear solution was dried over sodium sulfate, and filtered. Approximately 60% of the solvent was distilled off under vacuum and the resulting viscous oil was cooled to 10-15 ⁰C and mixed with diisopropyl ether (50 ml). The reaction mixture was stirred for 30-35 minutes and the resulting solid was filtered and dried at 40-60⁰C to obtain 1.75 g of the title compound. Example 18: Conversion of amorphous mitiglinide calcium into crystalline mitiglinide calcium A suspension of amorphous mitiglinide calcium in diisopropyl ether (30 ml) was stirred for 2 hours at 25- 30⁰C, filtered and dried under vacuum at 45-60⁰C to obtain crystalline form of mitiglinide calcium. Example 19: Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (2.5 g) in water (2.5 ml), aqueous ammonia solution (approx 25%, 4.0 ml) and acetonitrile (2.5 ml) at 10-15⁰C, calcium chloride (1.32 g) dissolved in demineralized water (15 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 25 ml) and acetone (2 x 5 ml) and dried at 45-50⁰C under vacuum to obtain 2.12 g of title compound having purity: 99.72 % by HPLC.

Example 20: Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (2.5 g) in water (2.5 ml), aqueous ammonia solution (approx 25%, 4.0 ml) and tetrahydrofuran (2.5 ml) at 10-15⁰C, calcium chloride (1.32 g) dissolved in demineralized water (15 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 25 ml) and acetone (2 x 5 ml) and dried at 45-50⁰C under vacuum to obtain 1.95 g of title compound having purity: 99.52 % by HPLC.

Example 21; Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (30.0 g) in water (300 ml), aqueous ammonia solution (approx 25%, 48 ml) and acetone (300 ml) at 10-15⁰C, calcium chloride (15.8 g) dissolved in demineralized water (180 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 300 ml) and acetone (2 x 60 ml) and dried at 45-50⁰C under vacuum to obtain 24.32 g of title compound having purity: 99.42 % by HPLC.

Example 22: Preparation of crystalline mitiglinide calcium

To a solution of mitiglinide (3.0 g) in water (30 ml), aqueous ammonia solution (approx 25%, 4.8 ml) and isopropyl alcohol (300 ml) at 10-15⁰C, calcium chloride (1.58 g) dissolved in demineralized water

(18 ml) was added. The mixture was stirred for 2 hours. The resulting precipitate was filtered, slurry washed with water (3 x 30 ml) and acetone (2 x 6 ml) and dried at 45-50⁰C under vacuum to obtain 1.92 g of title compound having purity: 99.65 % by HPLC.

Example 23: Preparation of (2S)-2-benzyWV-((lR)-l-benzyl-2-hydroxy-ethyl)-4-(hexahvdro- isoindolin-2-yl)-4-oxo-buryramide

To a solution of (2S)-2-benzyl-l-((4R)-4-benzyl-2-oxo-oxazolidin-3-yl)-4-(hexahydro-isoindolin-2-yl)- butane-l,4-dione (20.0 g) in tetrahydrofuran (140 ml), a solution of lithium hydroxide monohydrate

(3.43 g,) in demineralized water (40 ml) was added and the reaction mixture was refluxed for 4 hours till the completion of the reactions (monitored by thin layer chromatography). After the completion of the reaction, the reaction mixture was poured into demineralized water (100 ml) and extracted with ethyl acetate (2 x 80 ml). The combined organic layer was washed with water (80 ml) and dried over sodium sulphate. The solvent was evaporated under reduced pressure to give residue which was stirred in isopropyl alcohol at 0-5 ⁰C for 5 hours. The mixture was filtered and then dried at 40-45 ⁰C under vacuum to obtain 12.48 g of title compound having purity 99.77 % by HPLC. Melting point = 77 – 80⁰C.

PATENT

https://www.google.com/patents/CN102101838A?cl=en

Mitiglinide calcium (mitiglinide calcium), the chemical name (2S) _2_ benzyl _3_ (cis – hexahydro _2_ isoindolinyl-carbonyl) propionate dihydrate by Japanese pharmaceutical company developed Kissei ATP-dependent potassium channel blockers, 2004 for the first time in Japan for the treatment of type II diabetes.

Mitiglinide calcium is the second repaglinide, nateglinide after the first three columns MAG urea drugs, is a derivative of phenylalanine, which acts like mechanism sulfonylurea, but faster onset and the short half-life, is conducive to reducing postprandial blood glucose in diabetic patients, but also to avoid low blood sugar caused by continuous glucose, with “in vitro pancreas” reputation.

 In recent years, synthetic methods as described in patent application number: Patent 200510200127 9, the synthesis process first synthesized racemic (±) 2_-benzyl-3- (cis – hexahydro iso-indole-2. carbonyl) propionic acid, and then split to give (2S) -2- benzyl-3- (cis – hexahydro isoindole-2-carbonyl) propionic acid, not a lot of waste material along _ hexahydro isoindole, and Chiral separation is not high.

DISCLOSURE

The technical problem to be solved by the present invention is to provide a material savings along _ hexahydro isoindole, and preparation of a high degree of chiral separation.

To solve the above technical problem, the technical solution of the present invention is employed as a method for preparing mitiglinide calcium, comprising the steps of:

Step 1 Synthesis, benzylidene succinic acid

With stirring, was added sodium metal in absolute ethanol, under an inert gas, the solution was heated to reflux with stirring, reflux for 45 fly 0 minutes, under reflux before the dropwise addition of benzaldehyde, and then added dropwise diethyl succinate esters, reaction stirring was continued for 2 to 3 hours, slowly reducing the LC-Ms detection, the ratio of formaldehyde starting material benzene, cooled to room temperature, after use 5 (T55wt% aqueous solution of NaOH to adjust the PH San 13.0, and then heated at reflux;. Γ4 hours, cooled to at room temperature, keeping the reaction solution temperature <25 ° C, pH adjusted with concentrated hydrochloric San 2.0, filtration, recrystallization cold tetrahydrofuran, wherein the molar ratio of sodium metal with benzaldehyde and diethyl succinate is: 0.3 … ~ 0 5: 1 2~1 5: 1; Step 2 synthesis, benzyl butyl acid

The benzylidene succinic acid into the reactor, 10% Pd / C and ethanol, evacuated, and then replaced with hydrogen three times, introducing hydrogen, atmospheric hydrogenation reaction 12~15 hours, the reaction solution suction After the filtrate was evaporated to dryness under reduced pressure, the resulting solid was recrystallized from ethyl acetate, wherein the mass ratio of benzylidene succinic acid with 10% Pd / C is 1: 0 0 15 ^ 20.

3 Synthesis [0006] step, (S) -2- acid, benzyl butyl

Benzyl succinic acid dissolved in methanol was added dropwise with stirring (R) – a chiral amine, stirred at room temperature 2 hours wide, and the precipitated solid was filtered and the solid dispersed in water, under stirring 6 mol / mL hydrochloric acid adjusted ρΗ = 1 (Γ2.0, stirred for 30 minutes, the solid by suction filtration, dried, and wherein the benzyl succinic acid (R) – chiral amine molar ratio of 1: 0~2 5 2;…

Said (R) – a chiral amine (R) -I- phenylethylamine, (R) -I- naphthylethylamine or (R) -I- phenyl-2-p-amine;

4 (S) synthesis step, -2-benzyl succinic anhydride

Reactor, has added (S) -2- benzyl succinic acid and acetic anhydride, at 7 5,0 ° C reaction 1 to 2 hours, isopropyl ether low temperature crystallization after cooling, heavy with ethyl acetate crystallization, wherein (S) -2- molar ratio of benzyl succinic acid and acetic anhydride: 1: 7 · 0 to 7 · 5;

Step 5, (2S) -2- benzyl-3- (cis – hexahydro isoindole-2-carbonyl) propionic acid Synthesis

Stirring, S- benzyl succinic anhydride is dissolved in dichloromethane, control the internal temperature <0 V, a solution of cis _ hexahydro isoindole, dropping it, keeping the internal temperature at <0! : Continue stirring for 2 to 3 hours, the reaction in 2 (T25 ° C 10~15 hours, concentrated to give a pale yellow viscous material, wherein the (S) -2- benzyl succinic anhydride and cis – hexahydro isoindole molar ratio of 1: 2 (Γ2 5; step 6, mitiglinide calcium synthesis.

To the reactor was added (2S) -2- benzyl-3- (cis – hexahydro isoindole-2-carbonyl) propionic acid, water and concentrated ammonia, stir until completely dissolved, a solution of anhydrous calcium chloride aqueous solution, gradually precipitated white solid was added dropwise and then stirred at room temperature 12~ after 15 hours, suction filtered, the filter cake washed with water, dried to give a white solid, i.e. Mitiglinide calcium crude, obtained crude product with methanol and water (volume Than 0.5 4~0 5: 1) and recrystallized as a white solid Mitiglinide calcium;

Beneficial effects: The invention provides a method for preparing calcium Mitiglinide not only saves raw material cis – hexahydro isoindole, and chiral separation is high.

Embodiment 1

Step 1 Synthesis, benzylidene succinic acid

Under stirring, sodium metal (1.7 g, 0. 072 mol) was added absolute ethanol (50 mL), and under argon, the solution was heated to reflux with stirring, reflux for 50 minutes under reflux before the dropwise addition of benzene Formaldehyde (23 mL, 0. 183 mol), and then added dropwise diethyl succinate (50 mL, 0. 275 mol), stirring was continued for 2.5 hours the reaction, reducing the slow LC-Ms detection, the ratio of formaldehyde starting material benzene , was cooled to room temperature, with 55wt.% aqueous NaOH solution adjusting pH ≥ 13.0, and then heated at reflux for 3 hours, cooled to room temperature, the reaction solution temperature maintained <25 ° C, with concentrated hydrochloric pH≤2.0, leaching, cryogenic tetrahydrofuran recrystallization, yield = 81.3%;

Step 2 synthesis, benzyl butyl acid

The benzylidene succinic acid (23. 7 g, 0. 114 mol) into the reactor, then add 10% Pd / C (4. 7g) and anhydrous ethanol (300 mL), evacuated, then Hydrogen replacement three times, introducing hydrogen, hydrogenated at atmospheric pressure for 14 hours, the reaction solution after filtration, evaporated to dryness under reduced pressure, the resulting solid was recrystallized from ethyl acetate, yield: 98% 9; step 3, (S). -2-butyric acid benzyl

Benzyl succinic acid (31. 2 g, 0. 156 mol) was dissolved in methanol (500 mL), and added dropwise with stirring (R) -I- phenylethylamine (41.2 g, 0. 343 mol), room temperature stirred for 1.5 hours, the precipitated solid was filtered, the solid dispersion to water (100 mL) and stirred at with 6 mol / mL hydrochloric acid adjusted ρΗ = 1. (Γ2. 0, stirred for 30 minutes, the solid was suction filtered, and dried Yield 87. 3%; 4 (S) synthesis step, -2-benzyl succinic anhydride

Reactor, has added (S) -2- benzylbutyl acid (27. 8 g, 0. 132 mol) and acetic anhydride (88 mL, 0. 964 mol), at 7 5,0 ° C for 1 hours, cooled and added to isopropyl ether (150 mL) low temperature crystallization, after recrystallization from ethyl acetate, yield: 73% 9;.

Under – (hexahydro-isoindole-2-carbonyl cis) acid synthesis stirring S- benzyl succinic anhydride (12. 7 g, 0 Step 5, (2S) -2- benzyl-3. 067 mol) was dissolved in dichloromethane (250 mL), to control the internal temperature <0 ° C, a solution of cis – hexahydro isoindole (18. 5 g, 0 154 mol), the addition was complete, maintaining the internal temperature in <0! : Continue stirring for 2.5 hours, the reaction in 2 (T25 ° C 12 hours, concentrated to give a pale yellow viscous material, yield: 83 1%; Step 6 Synthesis Mitiglinide calcium.

To the reactor was added (2S) -2- benzyl-3- (cis – hexahydro isoindole-2-carbonyl) propionic acid (. 28. 7 g, 0 091 mol), water (150 mL), and concentrated aqueous ammonia (12 mL), stirring until completely dissolved, a solution of anhydrous calcium chloride (12. 1 g, 0.109 mol) water (100 mL) solution was gradually precipitated white solid was added dropwise at room temperature and then stirred for 13 End hours, filtration, washing the filter cake, and drying to give a white solid, crude Mitiglinide calcium, derived from crude methanol and water (volume ratio 0.5 4~0 5: 1) and recrystallized as a white solid MIG Chennai column calcium, yield: 87.3%.

 Second Embodiment

Example A similar experimental method steps 1 through 6 was carried out except in step 3, using (R) -1- naphthyl-amine (61. 4 g, 0. 359 mol) substituted (R) -I- phenylethylamine Other operating homogeneous reaction similar to this step of the synthesis yield: 87.3%.

Third Embodiment

Example A similar experimental method steps 1 through 6 was carried out except in step 3, using (R) -I- phenyl-2-p-tolyl-ethylamine (90. 2 g, 0. 374 mol) substituent (R ) -I- phenethylamine, other homogeneous reaction procedure similar to the synthesis yield of this step:. 83 4% ο

References

External links

• Elixir Pharmaceuticals – website of the U.S. rights holder for mitiglinide.

Mitiglinide

Systematic (IUPAC) name
(2S)-2-benzyl-4-[(3aR,7aS)-octahydro-2H-isoindol- 2-yl]-4-oxobutanoic acid
Clinical data
AHFS/Drugs.com	International Drug Names
Routes of administration	oral
Identifiers
CAS Number	145375-43-5
ATC code	A10BX08 (WHO)
PubChem	CID 121891
DrugBank	DB01252
ChemSpider	108739
UNII	D86I0XLB13
KEGG	D01854
ChEMBL	CHEMBL471498
Chemical data
Formula	C19H25NO3
Molar mass	315.41 g/mol

/////////207844-01-7, 145525-41-3, KAD-1229,  S-21403, MITIGLINIDE, Glufast, Kissei, 145375-43-5

Quality Control & MSDS

O=C(O)[C@@H](Cc1ccccc1)CC(=O)N3C[C@H]2CCCC[C@H]2C3

MF C22H20N4O2, MW 372.4

(S)-2-(5-(cyclopropylethynyl)-4-phenyl-1H-1,2,3-triazol-1-yl)-N-hydroxy-3-phenylpropanamide

1H NMR (500 MHz, d4-MeOD) 0.80 (2H, m), 0.98 (2H, m), 1.47 (1H, m), 3.51 (1H, dd, J = 11.2, 14.2 Hz), 3.71 (1H, dd, J = 3.9, 14.2 Hz), 5.49 (1H, dd, J = 3.9, 11.2 Hz), 6.96 (2H, m), 7.17-7.20 (3H, m), 7.37 (1H, t, J = 7.3 Hz), 7.43 (2H, t, J = 7.3 Hz), 7.99 (2H, d, J = 8.8 Hz);

13C NMR (100 MHz, d4-MeOD) 0.02, 8.55, 37.07, 60.83, 62.59, 109.09, 118.98, 125.9, 127.16, 128.55, 128.65, 128.71, 129.16, 130.07, 136.09, 147.10, 165.20;

HRMS calculated for C22H21N4O2 + (M+H): 373.1659, found: 373.1665.

PATENT

WO2014116962

https://www.google.com/patents/WO2014116962A1?cl=en

SAR. libraries were synthesized to investigate substitution about the triazole core. In some examples, compounds were synthesized using the synthetic routes shown in Fig. 2.

In one study, compound
was synthesized as outline in Scheme I.

Scheme I

PATENT

US153441899

https://patentscope.wipo.int/search/en/detail.jsf?docId=US153441899&recNum=1&office=&queryString=FP%3A%28Aaron+Beeler%29&prevFilter=&sortOption=Pub+Date+Desc&maxRec=8

 was synthesized as outline in Scheme I.

Paper

A novel, isoform-selective inhibitor of histone deacetylase 8 (HDAC8) has been discovered by the repurposing of a diverse compound collection. Medicinal chemistry optimization led to the identification of a highly potent (0.8 nM) and selective inhibitor of HDAC8.

Development of a Potent and Selective HDAC8 Inhibitor

http://pubs.acs.org/doi/abs/10.1021/acsmedchemlett.6b00239

file:///C:/Users/Inspiron/Downloads/ml6b00239_si_001.pdf

Department of Chemistry and Center for Molecular Discovery (BU-CMD), Boston University, 590 Commonwealth Avenue, Boston, Massachusetts 02215, United States

Center for Molecular Discovery (CMD) Director John Porco and members of the CMD lab team.

Aaron Beeler

Aaron Beeler received his Ph.D. in 2002 from Professor John Rimoldi’s laboratory in the Department of Medicinal Chemistry at the University of Mississippi. He then joined the Porco group as a postodoctoral fellow and subsequently the Center for Chemical Methodology and Library Development at Boston University, now the Center for Molecular Discovery. He was promoted to Assistant Director of the CMLD-BU in January 2005. In 2012 Aaron joined the Department of Chemistry as a tenure-track professor in medicinal chemistry.

Degrees and Positions

• B.S. Belmont University, Biology,
• Ph.D. University of Mississippi, Medicinal Chemistry

Research

The Beeler Research Group is truly multidisciplinary, combining organic chemistry, engineering, and biology to solve problems in medicinal chemistry. All of these elements are combined and directed toward significant problems in human health. The Beeler Group is addressing focused disease areas (e.g., schizophrenia, Parkinson’s, cystic fibrosis), as well as project areas with broader impact potential (e.g., new methods for discovery of small molecules with anti-cancer properties).

• Medicinal Chemistry: The goals of medicinal chemistry projects are to optimize small molecules in order to: a) develop a probe that may be utilized as a tool in biological studies; b) develop a lead molecule to facilitate future therapeutics; and c) utilize small molecules to enhance understanding of biological targets that are important for human health. These projects provide students with training in organic chemistry, medicinal chemistry, and focused biology. Projects are selected based on their chemistry and/or biology significance and potential for addressing challenging questions.
• Technology: One of the core components of the research in the Beeler Group is development of technologies and paradigms that facilitate rapid modification of complex scaffolds. These technologies enable optimization of biologically active lead compounds and identification of small molecule leads in biological systems. The projects focus on utilizing automation, miniaturization, and microfluidics to carry out chemical transformations. These projects are highly interdisciplinary with both chemistry and engineering components.
• Photochemistry: This area focuses on photochemical transformations toward the synthesis of natural products, natural product scaffolds, and other complex chemotypes of interest to medicinal chemistry and chemical biology. The foundation of these projects is utilizing microfluidics to enable photochemical reaction development.

Techniques & Resources

Students in the Beeler Research Group will have opportunities to learn a number of exciting research disciplines. Organic synthesis will be at the heart of every project. This will include targeted synthesis, methodology development, and medicinal chemistry. Through collaborations with biological researchers and/or research projects carried out within the Beeler Group, students will learn methods for biological assays, pharmacology, and target identification. Many projects will also include aspects of engineering that will provide opportunities for learning techniques such as microfabrication and microfluidics.

Opportunities

It is becoming evident that successful and impactful science is realized in collaborative interdisciplinary environments. The Beeler Research Group’s multidisciplinary nature and collaborative projects provides opportunities to learn areas of research outside of traditional chemistry.

What’s Next for Graduates of the Beeler Group?

Members of the Beeler Research Group will be positioned for a wide range of future endeavors.

• Undergraduates will be prepared to enter into graduate school for organic chemistry, chemical biology, or chemical engineering or to start careers in industry;
• Graduate students will have the foundation required for postdoctoral studies in organic synthesis or chemical biology as well as an industrial career in biotech or pharma;
• Postdoctoral associates will gain training and experience critical for both academic and industrial careers.

Assistant Professor
Office: SCI 484C
Laboratory: SCI 484A
Phone: 617.358.3487
Fax: 617-358-2847
beelera@bu.edu
Office Hours: by Appointment
Beeler Group Homepage
Google Scholar Page

Oscar J. Ingham below

John A. PORCO, JR  below

JAMES E. BRADNER, MD  above

Dana-Farber Cancer Institute

///////////epigenetic,  HDAC, HDAC8,  Histone deacetylase,  histone deacetylase 8,  triazole, PRECLINICAL, Department of Chemistry and Center for Molecular Discovery (BU-CMD), Boston University, 590 Commonwealth Avenue, Boston, Massachusetts 02215, United States, Oscar J. Ingham, Aaron Beeler

n1n(c(c(n1)c2ccccc2)C#CC3CC3)C(C(=O)NO)Cc4ccccc4

PF 4136309

PF4136309; PF 4136309; PF-4136309; PF04136309; PF4136309; PF-04136309; INCB8761; INCB 8761; INCB-8761

(S)-N-(2-(3-((4-hydroxy-4-(5-(pyrimidin-2-yl)pyridin-2-yl)cyclohexyl)amino)pyrrolidin-1-yl)-2-oxoethyl)-3-(trifluoromethyl)benzamide

N-[2-[(3S)-3-[[trans-4-Hydroxy-4-[5-(2-pyrimidinyl)-2-pyridinyl]cyclohexyl]amino]-1-pyrrolidinyl]-2-oxoethyl]-3-(trifluoromethyl)benzamide

N-[2-((3S)-3-[4-hydroxy-4-(4-pyrimidin-2-ylphenyl)cyclohexyl]aminopyrrolidin-1-yl)-2- oxoethyl]-3-(trifluoromethyl)benzamide

1341224-83-6
MF: C29H31F3N6O3
MW: 568.24097

CC chemokine receptor 2 (CCR2) antagonist

Pfizer Limited

Gary Burgess

PF-4136309, also known as INCB8761, is an orally available human chemokine receptor 2 (CCR2) antagonist with potential immunomodulating and antineoplastic activities. Upon oral administration, CCR2 antagonist PF-04136309 specifically binds to CCR2 and prevents binding of the endothelium-derived chemokine ligand CLL2 (monocyte chemoattractant protein-1 or MCP1) to its receptor CCR2, which may result in inhibition of CCR2 activation and signal transduction. This may inhibit inflammatory processes as well as angiogenesis, tumor cell migration, and tumor cell proliferation. The G-protein coupled receptor CCR2 is expressed on the surface of monocytes and macrophages, stimulates the migration and infiltration of these cell types, and plays an important role in inflammation, angiogenesis, and tumor cell migration and proliferation.

• Originator Pfizer
• Class Analgesics
• Mechanism of Action CCR2 receptor antagonists

Highest Development Phases

• Phase I/II Pancreatic cancer
• Discontinued Hepatic fibrosis; Pain

Most Recent Events

• 01 Apr 2016 Phase-I/II clinical trials in Pancreatic cancer (Combination therapy, First-line therapy, Metastatic disease) in USA (PO) (NCT02732938)
• 01 Dec 2015 Phase-I clinical trials in Pancreatic cancer (In volunteers) in Belgium (PO) (NCT02598206)
• 09 Nov 2015 Pfizer plans a phase I trial in Healthy volunteers in Belgium and USA (NCT02598206)

(S)-N-[2-(3-{trans-4-Hydroxy-4-[5-(pyrimidin-2-yl)pyridin-2-
yl]cyclohexylamino}pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl)benzamide

MS (M+H)+:569.2.

1H NMR (400 MHz, CD3OD): δ 9.57 – 9.45 (m, 1H), 8.94-8.84 (m, 2H), 8.82 –
8.72 (m, 1H), 8.27 – 8.19 (m, 1H), 8.15 (d, J = 7.8 Hz, 1H), 7.91 – 7.84 (m, 2H), 7.69
(dd, J = 7.8, 7.8 Hz, 1H), 7.46-7.39 (m, 1H), 4.29 – 4.12 (m, 2H), 3.87 (dd, J = 10.1, 6.4
Hz, 0.5H), 3.83 – 3.39 (m, 3.5H), 3.38 – 3.32 (m, 1H), 3.02 – 2.91 (m, 1H), 2.51 – 2.35
(m, 2H), 2.34 – 2.14 (m, 1H), 2.13 – 1.88 (m, 2.5H), 1.88 – 1.76 (m, 0.5H), 1.74 – 1.56
(m, 4H).

Anal. (C29H31F3N6O3): calcd C 61.24, H 5.50, N 14.79; found C 61.18, H 5.59,
N 14.87.

INTERMEDIATES

8-(5-Bromopyridin-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol

LC-MS (M+H)+: 316.1/314.1. 1H NMR (300 MHz,CDCl3): δ 8.60 (s, 1 H), 7.82 (d, 1 H), 7.38 (d, 1 H), 4.6 (s, 1 H), 4.0 (m, 4 H), 2.2 (m, 4
H), 1.7 (m, 4 H).

8-(5-Pyrimidin-2-ylpyridin-2-yl)-1,4-dioxaspiro[4.5]decan-8-ol

LC-MS (M+H)+: 314.2.

4-Hydroxy-4-(5-pyrimidin-2-ylpyridin-2-yl)cyclohexanone

MS
(M+H)+: 270.2.

tert-Butyl [(S)-1-({[3-(Trifluoromethyl)benzoyl]amino}acetyl)
pyrrolidin-3-yl]carbamate.

MS (M-Boc+H)+: 316.

(S)-N-{2-[3-Aminopyrrolidin-1-yl]-2-oxoethyl}-3-(trifluoromethyl)
benzamide hydrochloride

MS
(M+H)+: 316.

PATENT

WO 2012114223

https://www.google.com/patents/WO2012114223A1?cl=en

Example 35

Step A

8-(4-lodo-phenyl)-1 ,4-dioxa-spiro[4.5]decan-8-ol. To a solution of 1 ,4-diiodobenzene (16.5 g, 50 mmol) in THF (350 mL) at -78°C was added n-BuLi (2.5 M, 24 mL) over 1 hour. After stirred additional 30 minutes, a solution of 1 ,4-dioxa-spiro[4.5]decan-8-one (7.8 g, 50 mmol) in THF (30 mL) was added in and the resulting mixture was stirred for 3 hours. To the mixture was added TMSCI (5.4 g, 50 mmol) and the resulting mixture was allowed to warm to rt and stirred at rt for 18 hours. The reaction mixture was neutralized to pH 6.0, and extracted with ethyl acetate (3X 50 mL). The organic extracts were combined, washed with saline solution (2X 50 mL), dried over sodium sulfate, concentrated in vacuo. The residue was chromatographed on silica gel, eluting with hexane/ethyl acetate (95/5 to 100/0). The appropriate fractions were combined to give 8-(4-lodo-phenyl)-1 ,4-dioxa-spiro[4.5]decan-8-ol (12 g, 66.6%) with LCMS: 361 .2 (M+H⁺, 100%) and {[8-(4-iodophenyl)-1 ,4- dioxaspiro[4.5]dec-8-yl]oxy}(trimethyl)silane (6 g, 27%) with LCMS: 433.1 (M+H⁺, 100%). Step B

8-(4-pyrimidin-2-ylphenyl)-1 ,4-dioxaspiro[4.5]decan-8-ol. To a solution of 8-(4-iodo- phenyl)-1 ,4-dioxa-spiro[4.5]decan-8-ol (450.0 mg, 1.249 mmol) in THF (1.0 mL) at room temperature was added dropwise isopropylmagnesium chloride (2.0 M in THF, 1 .37 mL) and the reaction mixture was stirred at room temperature for 30 mins. To another flask charged with nickel acetylacetonate (20 mg, 0.06 mmol) and 1 ,3-bis(diphenylphosphino)-propane (26 mg, 0.062 mmol) suspened in THF (3 mL) under N2 was added 2-bromopyrimidine (199 mg, 1.25 mmol). The resulting mixture was stirred at room temperature until it is clear. The second mixture was transferred into the degassed Grignard solution prepared in step 1. The resulting mixture was stirred at room temperature overnight. The reaction mixture was diluted with EtOAc, quenched with water, washed with brine, dried overNa₂S0₄, and concentrated. The residue was columned on silica gel, eluted with hexane/EtOAc (2/1 ), to gave the desired compound (270 mg, 69%) as white solid. LCMS: 313.1 , (M+H, 100%). ¹H

NMR (CDCIs): δ 8.86 (d, 2H), 8.46 (dd, 2H), 7.71 (dd, 2H), 7.24 (t, 1 H), 4.05 (d, 4H), 2.30 (dt, 2H), 2.18 (dt, 2H), 1 .90 (m, 2H), 1 .78 (m, 2H).

Step C

4-Hydroxy-4-(4-pyrimidin-2-ylphenyl)cyclohexanone. The title compound was prepared by treating the ketal of step B with HCI in water following the procedure described in step B of Example 2. MS (M+H)⁺ 269.

Step D

N-[2-((3S)-3-[4-hydroxy-4-(4-pyrimidin-2-ylphenyl)cyclohexyl]aminopyrrolidin-1-yl)-2- oxoethyl]-3-(trifluoromethyl)benzamide bis(trifluoroacetate) (salt). To a 1-neck round-bottom flask charged with methylene chloride (1 ml.) was added 4-hydroxy-4-(4-pyrimidin-2- ylphenyl)cyclohexanone (50.0 mg, 0.186 mmol), N-2-[(3S)-3-aminopyrrolidin-1-yl]-2- oxoethyl-3-(trifluoromethyl)benzamide hydrochloride (65.5 mg, 0.186 mmol), and triethylamine (85.7 uL, 0.615 mmol). The resulting mixture was stirred at 25°C for 30 minutes, and to it was added sodium triacetoxyborohydride (62.4 mg, 0.28 mmol) in portion. The reaction mixture was stirring at rt overnight. The reaction was concentrated, and the residue was chromatographed on Si02, eluted with acetone/methanol (100% to 90%/10%) to give two fractions, which were further purified on prep-LCMS separately to afford F1 (24.2 mg ) and F2 (25.9 mg) as white powder in total 34% of the yield. LCMS: 568.2 (M+H, 100%)

Paper

Discovery of INCB8761/PF-4136309, a Potent, Selective, and Orally Bioavailable CCR2 Antagonist

Chu-Biao Xue*†, Anlai Wang†, Qi Han†, Yingxin Zhang†, Ganfeng Cao†, Hao Feng†, Taisheng Huang†,Changsheng Zheng†, Michael Xia†, Ke Zhang†, Lingquan Kong†, Joseph Glenn†, Rajan Anand†, David Meloni†,D. J. Robinson†, Lixin Shao†, Lou Storace†, Mei Li†, Robert O. Hughes‡, Rajesh Devraj‡, Philip A. Morton‡, D. Joseph Rogier‡, Maryanne Covington†, Peggy Scherle†, Sharon Diamond†, Tom Emm†, Swamy Yeleswaram†,Nancy Contel†, Kris Vaddi†, Robert Newton†, Greg Hollis†, and Brian Metcalf†

Incyte Corporation, Experimental Station E336, Wilmington, Delaware 19880, United States

Pfizer Global Research and Development, Chesterfield Parkway West, St. Louis, Missouri 63017, United States

ACS Med. Chem. Lett., 2011, 2 (12), pp 913–918

DOI: 10.1021/ml200199c, http://pubs.acs.org/doi/abs/10.1021/ml200199c

Tel: 302-498-6706. Fax: 302-425-2750. E-mail: cxue@incyte.com.

We report the discovery of a new (S)-3-aminopyrrolidine series of CCR2 antagonists. Structure–activity relationship studies on this new series led to the identification of 17 (INCB8761/PF-4136309) that exhibited potent CCR2 antagonistic activity, high selectivity, weak hERG activity, and an excellent in vitro and in vivo ADMET profile. INCB8761/PF-4136309 has entered human clinical trials.

HPLC

http://link.springer.com/article/10.1007/s10337-015-2860-8

A precise and sensitive LC method was developed and further validated for the determination of enantiomeric purity of (S)-N-[2-(3-{trans-4-hydroxy-4-[5-(pyrimidin-2-yl)pyridin-2-yl] cyclohexylamino} pyrrolidin-1-yl)-2-oxoethyl]-3-(trifluoromethyl) benzamide (PF-04136309). Baseline separation with a resolution higher than 1.8 was accomplished within 40 min using a CHIRALPAK AD (250 × 4.6 mm; particle size 5 μm) column, with n-hexane:2-propanol (70:30v/v) as mobile phase at a flow rate of 1 mL min⁻¹. The eluted analytes were subsequently detected with a UV detector at 260 nm. The effects of mobile phase components and temperature on enantiomeric selectivity as well as the resolution of enantiomers were thoroughly investigated. The calibration curves were plotted within a concentration range between 0.01 and 1 mg mL⁻¹ (n = 9), and recoveries between 98.17 and 101.28 % were obtained, with relative standard deviation (RSD) lower than 1.44 %. The LOD and LOQ for PF-04136309 were 3.59 and 11.54 μg mL⁻¹ and for its enantiomer were 3.39 and 11.28 μg mL⁻¹, respectively. The developed method was demonstrated to be accurate, robust and sensitive for the determination of enantiomeric purity of PF-04136309, especially for the analysis of bulk samples.

REFERENCES

1: Xue CB, Wang A, Han Q, Zhang Y, Cao G, Feng H, Huang T, Zheng C, Xia M, Zhang K, Kong L, Glenn J, Anand R, Meloni D, Robinson DJ, Shao L, Storace L, Li M, Hughes RO, Devraj R, Morton PA, Rogier DJ, Covington M, Scherle P, Diamond S, Emm T, Yeleswaram S, Contel N, Vaddi K, Newton R, Hollis G, Metcalf B. Discovery of INCB8761/PF-4136309, a Potent, Selective, and Orally Bioavailable CCR2 Antagonist. ACS Med Chem Lett. 2011 Oct 5;2(12):913-8. doi: 10.1021/ml200199c. eCollection 2011 Dec 8. PubMed PMID: 24900280; PubMed Central PMCID: PMC4018168.

http://www.pfizer.com/files/news/asco/ASCO2016_PipelineFactSheet_CCR2.pdf

//////1341224-83-6, PF 4136309, PF4136309,  PF 4136309, PF-4136309, PF04136309, PF4136309, PF-04136309, INCB8761, INCB 8761, INCB-8761, PFIZER, PHASE 2

O=C(NCC(N1C[C@@H](NC2CCC(C3=NC=C(C4=NC=CC=N4)C=C3)(O)CC2)CC1)=O)C5=CC=CC(C(F)(F)F)=C5

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Evofosfamide, HAP-302 , TH-302, TH 302

эвофосфамид ,  إيفوفوسفاميد ,  艾伏磷酰胺 ,

• Molecular Formula C₉H₁₆Br₂N₅O₄P
• Average mass 449.036 Da

(1-Methyl-2-nitro-1H-imidazol-5-yl)methyl N,N’-bis(2-bromoethyl)phosphorodiamidate

TH-302 is a nitroimidazole-linked prodrug of a brominated derivative of an isophosphoramide mustard previously used in cancer drugs

• Originator Threshold Pharmaceuticals
• Developer Merck KGaA; Threshold Pharmaceuticals
• Class Antineoplastics; Nitroimidazoles; Phosphoramide mustards; Small molecules
• Mechanism of Action Alkylating agents

• Orphan Drug Status Yes – Soft tissue sarcoma; Pancreatic cancer
• On Fast track Pancreatic cancer; Soft tissue sarcoma

• Suspended Glioblastoma; Leukaemia; Malignant melanoma; Multiple myeloma; Non-small cell lung cancer; Solid tumours
• Discontinued Pancreatic cancer; Soft tissue sarcoma

Most Recent Events

• 01 Aug 2016 Threshold plans a clinical trial for Solid tumours
• 01 Aug 2016 Threshold announces intention to submit NDA to the Pharmaceuticals and Medical Device Agency in Japan
• 16 Jun 2016 Merck KGaA terminates a phase II trial in Soft tissue sarcoma (Combination therapy, Inoperable/Unresectable, Metastatic disease, Late-stage disease) in Japan (IV) due to negative results from the phase III SARC021 trial (NCT02255110)

Evofosfamide (first disclosed in WO2007002931), useful for treating cancer.

Threshold Pharmaceuticals and licensee Merck Serono are codeveloping evofosfamide, the lead in a series of topoisomerase II-inhibiting hypoxia-activated prodrugs and a 2-nitroimidazole-triggered bromo analog of ifosfamide, for treating cancer, primarily soft tissue sarcoma and pancreatic cancer (phase 3 clinical, as of April 2015).

In November 2014, the FDA granted Fast Track designation to the drug for the treatment of previously untreated patients with metastatic or locally advanced unresectable soft tissue sarcoma.

Evofosfamide (INN,^[1] USAN;^[2] formerly known as TH-302) is an investigational hypoxia-activated prodrug that is in clinical development for cancer treatment. The prodrug is activated only at very low levels of oxygen (hypoxia). Such levels are common in human solid tumors, a phenomenon known as tumor hypoxia.^[3]

Evofosfamide is being evaluated in clinical trials for the treatment of multiple tumor types as a monotherapy and in combination with chemotherapeutic agents and other targeted cancer drugs.

Dec 2015 : two Phase 3 trials fail, Merck will not apply for a license

Collaboration

Evofosfamide was developed by Threshold Pharmaceuticals Inc. In 2012, Threshold signed a global license and co-development agreement for evofosfamide with Merck KGaA, Darmstadt, Germany (EMD Serono Inc. in the US and Canada), which includes an option for Threshold to co-commercialize evofosfamide in the United States. Threshold is responsible for the development of evofosfamide in the soft tissue sarcoma indication in the United States. In all other cancer indications, Threshold and Merck KGaA are developing evofosfamide together.^[4] From 2012 to 2013, Merck KGaA paid 110 million US$ for upfront payment and milestone payments to Threshold. Additionally, Merck KGaA covers 70% of all evofosfamide development expenses.^[5]

Mechanism of prodrug activation and Mechanism of action (MOA) of the released drug[edit]

Evofosfamide is a 2-nitroimidazole prodrug of the cytotoxin bromo-isophosphoramide mustard (Br-IPM). Evofosfamide is activated by a process that involves a 1-electron (1 e⁻) reduction mediated by ubiquitous cellular reductases, such as the NADPH cytochrome P450, to generate a radical anion prodrug:

• A) In the presence of oxygen (normoxia) the radical anion prodrug reacts rapidly with oxygen to generate the original prodrug and superoxide. Therefore, evofosfamide is relatively inert under normal oxygen conditions, remaining intact as a prodrug.
• B) When exposed to severe hypoxic conditions (< 0.5% O₂; hypoxic zones in many tumors), however, the radical anion undergoes irreversible fragmentation, releasing the active drug Br-IPM and an azole derivative. The released cytotoxin Br-IPM alkylates DNA, inducing intrastrand and interstrand crosslinks.^[6]

Evofosfamide is essentially inactive under normal oxygen levels. In areas of hypoxia, evofosfamide becomes activated and converts to an alkylating cytotoxic agent resulting in DNA cross-linking. This renders cells unable to replicable their DNA and divide, leading to apoptosis. This investigational therapeutic approach of targeting the cytotoxin to hypoxic zones in tumors may cause less broad systemic toxicity that is seen with untargeted cytotoxic chemotherapies.^[7]

The activation of evofosfamide to the active drug Br-IPM and the mechanism of action (MOA) via cross-linking of DNA is shown schematically below:

Drug development history

Phosphorodiamidate-based, DNA-crosslinking, bis-alkylator mustards have long been used successfully in cancer chemotherapy and include e.g. the prodrugs ifosfamide andcyclophosphamide. To demonstrate that known drugs of proven efficacy could serve as the basis of efficacious hypoxia-activated prodrugs, the 2-nitroimidizole HAP of the active phosphoramidate bis-alkylator derived from ifosfamide was synthesized. The resulting compound, TH-281, had a high HCR (hypoxia cytotoxicity ratio), a quantitative assessment of its hypoxia selectivity. Subsequent structure-activity relationship (SAR) studies showed that replacement of the chlorines in the alkylator portion of the prodrug with bromines improved potency about 10-fold. The resulting, final compound is evofosfamide (TH-302).^[8]

Synthesis

Evofosfamide can be synthesized in 7 steps.^[9][10]

1. CPhI.cn: Synthetic routes to explore anti-pancreatic cancer drug Evofosfamide, 22 Jan 2015
2.  Synthetic route Reference: International patent application WO2007002931A2

Formulation

The evofosfamide drug product formulation used until 2011 was a lyophilized powder. The current drug product formulation is a sterile liquid containing ethanol,dimethylacetamide and polysorbate 80. For intravenous infusion, the evofosfamide drug product is diluted in 5% dextrose in WFI.^[11]

Diluted evofosfamide formulation (100 mg/ml evofosfamide, 70% ethanol, 25% dimethylacetamide and 5% polysorbate 80; diluted to 4% v/v in 5% dextrose or 0.9% NaCl) can cause leaching of DEHP from infusion bags containing PVC plastic.^[12]

Clinical trials

Overview and results

Evofosfamide (TH-302) is currently being evaluated in clinical studies as a monotherapy and in combination with chemotherapy agents and other targeted cancer drugs. The indications are a broad spectrum of solid tumor types and blood cancers.

Evofosfamide clinical trials (as of 21 November 2014)^[13] sorted by (Estimated) Primary Completion Date:^[14]

Both, evofosfamide and ifosfamide have been investigated in combination with doxorubicin in patients with advanced soft tissue sarcoma. The study TH-CR-403 is a single arm trial investigating evofosfamide in combination with doxorubicin.^[35] The study EORTC 62012 compares doxorubicin with doxorubicin plus ifosfamide.^[36] Doxorubicin and ifosfamide are generic products sold by many manufacturers.Soft tissue sarcoma

The indirect comparison of both studies shows comparable hematologic toxicity and efficacy profiles of evofosfamide and ifosfamide in combination with doxorubicin. However, a longer overall survival of patients treated with evofosfamide/doxorubicin (TH-CR-403) trial was observed. The reason for this increase is probably the increased number of patients with certain sarcoma subtypes in the evofosfamide/doxorubicin TH-CR-403 trial, see table below.

However, in the Phase 3 TH-CR-406/SARC021 study (conducted in collaboration with the Sarcoma Alliance for Research through Collaboration (SARC)), patients with locally advanced unresectable or metastatic soft tissue sarcoma treated with evofosfamide in combination with doxorubicin did not demonstrate a statistically significant improvement in OS compared with doxorubicin alone (HR: 1.06; 95% CI: 0.88 – 1.29).

Metastatic pancreatic cancer

Both, evofosfamide and protein-bound paclitaxel (nab-paclitaxel) have been investigated in combination with gemcitabine in patients with metastatic pancreatic cancer. The study TH-CR-404 compares gemcitabine with gemcitabine plus evofosfamide.^[39] The study CA046 compares gemcitabine with gemcitabine plus nab-paclitaxel.^[40] Gemcitabine is a generic product sold by many manufacturers.

The indirect comparison of both studies shows comparable efficacy profiles of evofosfamide and nab-paclitaxel in combination with gemcitabine. However, the hematologic toxicity is increased in patients treated with evofosfamide/gemcitabine (TH-CR-404 trial), see table below.

In the Phase 3 MAESTRO study, patients with previously untreated, locally advanced unresectable or metastatic pancreatic adenocarcinoma treated with evofosfamide in combination with gemcitabine did not demonstrate a statistically significant improvement in overall survival (OS) compared with gemcitabine plus placebo (hazard ratio [HR]: 0.84; 95% confidence interval [CI]: 0.71 – 1.01; p=0.0589).

Drug development risks

Risks published in the quarterly/annual reports of Threshold and Merck KGaA that could affect the further development of evofosfamide (TH-302):

Risks related to the formulation

The evofosfamide formulation that Threshold and Merck KGaA are using in the clinical trials was changed in 2011^[43] to address issues with storage and handling requirements that were not suitable for a commercial product. Additional testing is ongoing to verify if the new formulation is suitable for a commercial product. If this new formulation is also not suitable for a commercial product another formulation has to be developed and some or all respective clinical phase 3 trials may be required to be repeated which could delay the regulatory approvals.^[44]

Risks related to reimbursement

Even if Threshold/Merck KGaA succeed in obtaining regulatory approvals and bringing evofosfamide to the market, the amount reimbursed for evofosfamide may be insufficient and could adversely affect the profitability of both companies. Obtaining reimbursement for evofosfamide from third-party and governmental payors depend upon a number of factors, e.g. effectiveness of the drug, suitable storage and handling requirements of the drug and advantages over alternative treatments.

There could be the case that the data generated in the clinical trials are sufficient to obtain regulatory approvals for evofosfamide but the use of evofosfamide has a limited benefit for the third-party and governmental payors. In this case Threshold/Merck KGaA could be forced to provide supporting scientific, clinical and cost effectiveness data for the use of evofosfamide to each payor. Threshold/Merck KGaA may not be able to provide data sufficient to obtain reimbursement.^[45]

Risks related to competition

Each cancer indication has a number of established medical therapies with which evofosfamide will compete, for example:

• If approved for commercial sale for pancreatic cancer, evofosfamide would compete with gemcitabine (Gemzar), marketed by Eli Lilly and Company; erlotinib (Tarceva), marketed by Genentech and Astellas Oncology; protein-bound paclitaxel (Abraxane), marketed by Celgene; and FOLFIRINOX, which is a combination of generic products that are sold individually by many manufacturers.
• If approved for commercial sale for soft tissue sarcoma, evofosfamide could potentially compete with doxorubicin or the combination of doxorubicin and ifosfamide, generic products sold by many manufacturers.^[46]

Risks related to manufacture and supply

Threshold relies on third-party contract manufacturers for the manufacture of evofosfamide to meet its and Merck KGaA’s clinical supply needs. Any inability of the third-party contract manufacturers to produce adequate quantities could adversely affect the clinical development and commercialization of evofosfamide. Furthermore, Threshold has no long-term supply agreements with any of these contract manufacturers and additional agreements for more supplies of evofosfamide will be needed to complete the clinical development and/or commercialize it. In this regard, Merck KGaA has to enter into agreements for additional supplies or develop such capability itself. The clinical programs and the potential commercialization of evofosfamide could be delayed if Merck KGaA is unable to secure the supply.^[47]

History

CLIP

CLIP

http://pubs.rsc.org/en/content/articlelanding/2015/qo/c5qo00211g/unauth#!divAbstract

http://www.rsc.org/suppdata/c5/qo/c5qo00211g/c5qo00211g1.pdf

Hypoxia, regions of low oxygen, occurs in a range of biological environments, and is involved in human diseases, most notably solid tumours. Exploiting the physiological differences arising from low oxygen conditions provides an opportunity for development of targeted therapies, through the use of bioreductive prodrugs, which are selectively activated in hypoxia. Herein, we describe an improved method for synthesising the most widely used bioreductive group, 2-nitroimidazole. The improved method is applied to an efficient synthesis of the anti-cancer drug Evofosfamide (TH-302), which is currently in Phase III clinical trials for treatment of a range of cancers.

(1-Methyl-2-nitro-1H-imidazol-5-yl)-N,N–bis(2-bromoethyl) phosphordiamidate (TH- 302)

The residue was then purified by semi-preparative HPLC on a Phenomenex Luna (C18(2), 10 µm, 250 × 10 mm) column, eluting with H2O and methanol (50 – 70% methanol over 10 min, then 1 min wash with methanol, 5 mL/min flow rate) to afford TH-302 as a yellow gum: vmax (solid) cm-1 : 3212 (br), 1489 (m), 1350 (m), 1105 (m), 1004 (s); δH (DMSO-D6, 400 MHz) 7.25 (1H, s, CH), 5.10–4.90 (2H, m, NHCH2CH2Br), 4.98 (2H, d, J 7.8, CH2O), 3.94 (3H, s, CH3), 3.42 (4H, t, J 7.0, NHCH2CH2Br), 3.11 (4H, dt, J 9.8, 7.2, NHCH2CH2Br); δC (DMSO-D6, 126 MHz) 146.1, 134.2 (d, J 7.5, OCH2CN), 128.2, 55.6 (d, J 4.6, CH2O), 42.7, 34.2 (d, J 26.4, CH2Br), 34.1; δP (DMSO-D6, 202 MHz) 15.4; HRMS m/z (ESI− ) [found; (M-H)− 447.9216, C9H16 79Br81BrN5O4P requires (M-H)− 447.9213]; m/z (ESI+ ) 448.0 ([M-H]− , 60%, [C9H15 79Br81BrN5O4P] − ), 493.9 ([M+formate] − , 100%, [C10H17 79Br81BrN5O6P] − ). These data are in good agreement with the literature values.4

4 J.-X. Duan, H. Jiao, J. Kaizerman, T. Stanton, J. W. Evans, L. Lan, G. Lorente, M. Banica, D. Jung, J. Wang, H. Ma, X. Li, Z. Yang, R. M. Hoffman, W. S. Ammons, C. P. Hart and M. Matteucci, J. Med. Chem., 2008, 51, 2412–2420.

J. Med. Chem., 2008, 51, 2412–2420/……………….1-Methyl-2-nitro-1H-imidazol-5-yl)methyl N,N-bis(2-bromoethyl)
phosphordiami-date (3b). Compound 3b was synthesized by a procedure similar to that described for 3a and obtained as an off-white solid in 47.6% yield.

1H NMR (DMSO-d6) δ: 7.22 (s, 1H), 5.10–5.00 (m, 2H), 4.97 (d, J ) 7.6 Hz, 2H), 3.94 (s, 3H), 3.42 (t, J ) 7.2 Hz, 4H), and 3.00–3.20 (m, 4H).

13C NMR (DMSOd6)δ: 146.04, 134.16 (d, J ) 32 Hz), 128.17, 55.64, 42.70, 34.33,and 34.11 (d, J ) 17.2 Hz).

31P NMR (DMSO-d6) δ: -11.25.
HRMS: Calcd for C9H16N5O4PBr2, 446.9307; found, 446.9294.

CLIP

Synthesis Route reference WO2007002931A2

Med J.. Chem. 2008, 51, 2412-2420

From compound S-1 starting aminoacyl protection is S-2 , a suspension of NaH grab α -proton, offensive, ethyl, acidification, introduction of an aldehyde group, S-3followed by condensation with the amino nitrile, off N- acyl ring closure, migration rearrangement amino imidazole compound S-. 8 , the amino and sodium nitrite into a diazonium salt, raising the temperature, nitrite anion nucleophilic attack diazonium salt obtained nitro compound S-9, under alkaline conditions ester hydrolysis gives acid S-10 , followed by NEt3 under the action of isobutyl chloroformate and the reaction mixed anhydride formed by of NaBH ₄ reduction to give the alcohol S-. 11 , [use of NaBH ₄ reduction of the carboxyl group is another way and the I ₂ / of NaBH ₄ ] , to give S-11 later, the DIAD / PPh3 ₃ under the action via Mitsunobu linking two fragments obtained reaction Evofosfamide

.

PATENT

http://www.google.co.in/patents/WO2015051921A1?cl=en

EXAMPLE 1

1

N-Formylsarcosine ethyl ester 1 (1 ,85 kg) was dissolved in toluene (3,9 kg) and ethyl formate (3,28 kg) and cooled to 10 °C. A 20 wt-% solution of potassium tert-butoxide (1 ,84 kg) in tetrahydrofuran (7,4 kg) was added and stirring was continued for 3h. The reaction mixture was extracted 2x with a solution of sodium chloride in water (10 wt-%) and the combined water extracts were washed lx with toluene.

Aqueous hydrogen chloride (25% wt-%; 5,62 kg) was added to the aqueous solution, followed by ethylene glycol (2,36 kg). The reaction mixture was heated to 55-60 °C for lh before only the organic solvent residues were distilled off under vacuum.

Aqueous Cyanamide (50 wt-%, 2,16 kg) was then added at 20 °C, followed by sodium acetate (3,04 kg). The resulting reaction mixture was heated to 85-90 °C for 2h and cooled to 0-5 °C before a pH of ~ 8-9 was adjusted via addition of aqueous sodium hydroxide (32% wt-%; 4,1 kg). Compound 3 (1,66 kg; 75%) was isolated after filtration and washing with water.

Ή-NMR (400 MHz, d6-DMSO): δ= 1,24 (3H, t, J= 7,1 Hz); 3,53 (3H, s); 4,16 (2H, q, J= 7,0 Hz) ; 6,15 (s, 2 H); 7,28 (s, 1H).

HPLC (R_t = 7,7 min): 97,9% (a/a).

HPLC data was obtained using Agilent 1100 series HPLC from agilent technologies using an Column: YMC-Triart CI 8 3μ, 100 x 4,6 mm Solvent A: 950 ml of ammonium acetate/acetic acid buffer at pH = 6 + 50 ml acetonitril; Solvent B: 200 ml of ammonium acetate/acetic acid buffer at pH = 6 + 800 ml acetonitril; Flow: 1,5 ml/min; Gradient: 0 min: 5 % B, 2 min: 5 % B, 7 min: 20 % B, 17 min: 85% B, 17, 1 min: 5% B, 22 min: 5% B.

PATENT

WO2007002931

http://www.google.com/patents/WO2007002931A2?cl=en

Example 8

Synthesis of Compounds 25, 26 [0380] To a solution of 2-bromoethylammmonium bromide (19.4 g) in DCM (90 mL) at – 1O⁰C was added a solution OfPOCl₃ (2.3 mL) in DCM (4 mL) followed by addition of a solution of TEA (14.1 mL) in DCM (25 mL). The reaction mixture was filtered, the filtrate concentrated to ca. 30% of the original volume and filtered. The residue was washed with DCM (3×25 mL) and the combined DCM portions concentrated to yield a solid to which a mixture of THF (6 mL) and water (8 mL) was added. THF was removed in a rotary evaporator, the resulting solution chilled overnight in a fridge. The precipitate obtained was filtered, washed with water (10 mL) and ether (30 mL), and dryed in vacuo to yield 2.1 g of:

Isophosphoramide mustard

can be synthesized employing the method provided in Example 8, substituting 2- bromoethylammmonium bromide with 2-chloroethylammmonium chloride. Synthesis of Isophosphoramide mustard has been described (see for example Wiessler et al., supra).

The phosphoramidate alkylator toxin:

was transformed into compounds 24 and 25, employing the method provided in Example 6 and the appropriate Trigger-OH.

Example 25

Synthesis of l-N-methyl-2-nitroimidazole-5-carboxylis acid

A suspension of the nitro ester (39.2 g, 196.9 rnmol) in IN NaOH (600 mL) and water (200 mL) was stirred at rt for about 20 h to give a clear light brown solution. The pH of the reaction mixture was adjusted to about 1 by addition of cone. HCl and the reaction mixture extracted with EA (5 x 150 mL). The combined ethyl acetate layers were dried over MgS O4 and concentrated to yield l-N-methyl-2-nitroimidazole-5-carboxylis acid (“nitro acid”) as a light brown solid (32.2 g, 95%). Example 26

Synthesis of l-N-methyl-2-nitroimidazole-5-carboxylis acid

A mixture of the nitro acid (30.82 g, 180.23 mmol) and triethylamine (140 niL, 285 mmol) in anhydrous THF (360 mL) was stirred while the reaction mixture was cooled in a dry ice-acetonitrile bath (temperature < -20 ⁰C). Isobutyl chloroformate (37.8 mL, 288 mmol) was added drop wise to this cooled reaction mixture during a period of 10 min and stirred for 1 h followed by the addition of sodium borohydride (36 g, 947 mmol) and dropwise addition of water during a period of 1 h while maintaining a temperature around or less than O⁰C. The reaction mixture was warmed up to O⁰C. The solid was filtered off and washed with THF. The combined THF portions were evaporated to yield l-N-methyl-2- nitroimidazole-5-methanol as an orange solid (25 g) which was recrystallized from ethyl acetate.

PATENT

WO-2015051921

EXAMPLE 1

1

N-Formylsarcosine ethyl ester 1 (1 ,85 kg) was dissolved in toluene (3,9 kg) and ethyl formate (3,28 kg) and cooled to 10 °C. A 20 wt-% solution of potassium tert-butoxide (1 ,84 kg) in tetrahydrofuran (7,4 kg) was added and stirring was continued for 3h. The reaction mixture was extracted 2x with a solution of sodium chloride in water (10 wt-%) and the combined water extracts were washed lx with toluene.

Aqueous hydrogen chloride (25% wt-%; 5,62 kg) was added to the aqueous solution, followed by ethylene glycol (2,36 kg). The reaction mixture was heated to 55-60 °C for lh before only the organic solvent residues were distilled off under vacuum.

Aqueous Cyanamide (50 wt-%, 2,16 kg) was then added at 20 °C, followed by sodium acetate (3,04 kg). The resulting reaction mixture was heated to 85-90 °C for 2h and cooled to 0-5 °C before a pH of ~ 8-9 was adjusted via addition of aqueous sodium hydroxide (32% wt-%; 4,1 kg). Compound 3 (1,66 kg; 75%) was isolated after filtration and washing with water.

Ή-NMR (400 MHz, d6-DMSO): δ= 1,24 (3H, t, J= 7,1 Hz); 3,53 (3H, s); 4,16 (2H, q, J= 7,0 Hz) ; 6,15 (s, 2 H); 7,28 (s, 1H).

HPLC (R_t = 7,7 min): 97,9% (a/a).

PATENT

WO 2016011195

http://google.com/patents/WO2016011195A1?cl=en

Figure 1 provides the differential scanning calorimetry (DSC) data of crystalline solid form A of TH-302.

Figure 2 shows the 1H-NMR of crystalline solid form A of TH-302.

Figure 5 shows the Raman Spectra of TH-302 (Form A)

Scheme 1 illustrates a method of preparing TH-302.

Scheme 1: Process for the Preparation of TH-302

NaOH (RGT)

Step 1. Imidazole Purified water (SLV)

Carboxylic Acid IPC: NMT 1.0% SM by HPLC

HCI (RGT)

IPC: pH 1.0 ± 0.5

IPC: NMT 1.0% water by KF

TH-302

MW = 449.0

SM = Starting Material INT = Intermediate IPC = In-process Control RGT = Reagent SLV = Solvent MW = Molecular Weight LOD = Loss on drying NMT = Not more than NLT = Not less than

TH-302 can be prepared by hydro lyzing (l-methyl-2-nitro-lH-imidazol-5-yl) ethyl ester above for example under aqueous conditions with a suitable base catalyst (e.g. NaOH in water at room temperature). The imidazole carboxylic acid prepared by this method can be used without further purification. However, it has been found that treating the dried crude intermediate product with a solvent such as acetonitrile, ethyl acetate, n-heptane, acetone, dimethylacetamide, dimethylformamide, 1, 4-dioxane, ethylene glycol, 2-propanol, 1-propanol, tetrahydrofuran (1 : 10 w/v) or combinations thereof in a vessel with heating, followed by cooling and filtration through a filtration aid with acetone decreased the number and levels of impurities in the product. The number and levels of impurities could be further reduced by treating the dried crude product with water (1 :5.0 w/v) in a vessel with heating followed by cooling and filtration through a filtration aid with water.

The carboxylic acid of the imidazole can then be reduced using an excess of a suitable reducing agent (e.g. sodium borohydride in an appropriate solvent, typically aqueous. The reaction is exothermic (i.e. potentially explosive) releasing borane and hydrogen gases over several hours. It was determined that the oxygen balance of the product imidazole alcohol is about 106.9, which suggests a high propensity for rapid decomposition. It has been found that using NaOH, for example 0.01M NaOH followed by quenching the reaction with an acid. Non-limiting examples of acids include, but are not limited to water, acetic acid, hydrobromic acid, hydrochloric acid, sodium hydrogen phosphate, sulfuric acid, citric acid, carbonic acid, phosphoric acid, oxalic acid, boric acid and combinations thereof. In some embodiments, the acid may diluted with a solvent, such as water and/or tetrahydrofuran. In some embodiments, acetic acid or hydrochloric acid provide a better safety profile, presumably because it is easier to control the temperature during the addition of the reducing agent and the excess reducing agent is destroyed after the reaction is complete. This also results in improved yields and fewer impurities, presumably due to reduced impurities from the reducing agent and decomposition of the product. Using this process, greater than 98.5% purity could be achieved for this intermediate. The formation of ether linkage can be accomplished by treating the product imidazole alcohol with solution of N,N’-Bis(2-bromoethyl)phosphorodiamidic acid (Bromo IPM), a trisubstituted phosphine and diisopropyl azodicarboxylate in tetrahydrofuran at room temperature to afford TH-302. It has been found that by recrystallizing the product from a solvents listed in the examples, one could avoid further purfication by column chromatography, which allowed for both reduced solvent use especially on larger scales.

Scheme 2 illustrates an alternative method of preparing TH-302.

Scheme 2: Process for the Preparation of TH-302

(SM)

ethylamine mide (SM) 04.9 ) SLV) , RGT) ter by KF

NT)

MW = 449.0

Example 1: Synthesis of TH-302

Step 1 – Preparation intermediate imidazole carboxylic

I T)

Crude imidazole carboxylic acid ethyl ester (1 : 1.0 w/w) was taken in water (1 : 10.0 w/v) at 25± 5°C and cooled to 17± 3°C. A 2.5 N sodium hydroxide solution (10 V) was added slowly at 17±3°C. The reaction mass was warmed to 25±5°C and monitored by HPLC. After the completion of reaction, the reaction mass was cooled to 3±2°C and pH of the reaction mass adjusted to 1=1=0.5 using 6 M HC1 at 3±2°C. The reaction mass was then warmed to 25±5°C and extracted with ethyl acetate (3 x 10 V). The combined organic layers

were washed with water (1 x 10 V) followed by brine (1 x 10 V). The organic layer was dried over sodium sulfate (3 w/w), filtered over Celite and concentrated. n-Heptane (1.0 w/v) was added and the the reaction mixture was concentrated below 45°C to 2.0 w/v. The reaction mass was cooled to 0±5°C. The solid was filtered, and the bed was washed with n-heptane (1 x 0.5 w/v) and dried at 35±5°C. In a vessel, acetone (1 : 10 w/v) was added. Dry crude imidazole carboxylic acid (ICA) from 1.12 was added to the acetone. The mixture was warmed to 45±5°C and was stirred for 30 minutes. The mass was cooled to 28±3°C and filtered through a Celite bed. The filter bed was washed with 1 : 1.0 w/v of acetone. Water (1 :5.0 w/v) was added to the filtrate and the mixture was concentrated. The concentrated mass was cooled to 5±5°C and stirred for 30 minutes. The material was filtered and the solid was washed 2 x 1 : 1.0 w/v of water at 3±2°C. The product was dried for 2 hours at 25±5°C and then at 45±5°C. As can be seen below, the number and levels of impurities are decreased.

Table I: Purity and Impurity Profile Comparison of Typical Crude ICA and Purified

ICA

Imidazole alcohol:

CI^Oi-Bu

T

o

Imidazole carboxylic acid (1.0 w/w) was taken in tetrahydrofuran (10 w/v) under nitrogen atmosphere at 25±5°C. The reaction mass was cooled to -15±5°C. Triethylamine (1 : 1.23 w/v) was added slowly over a period of 1 hour maintaining the temperature at – 15±5°C. The reaction mass was stirred at -15±5°C for 15-20 min. Isobutylchloroformate (1 : 1.14 w/v) was added slowly over a period of 1 hour maintaining the temperature at – 15±5°C. The reaction mass was stirred at -15±5°C for 30-40 min. A solution of sodium borohydride (1 : 1.15 w/w) in 0.01 M aqueous sodium hydroxide (2.2 w/v) was divided into 6 lots and added to the above reaction mass while maintaining the temperature of the reaction mass between 0±10°C for 40-60 min for each lot. The reaction mass was warmed to 25±5°C and stirred until imidazole carboxylic acid content < 5.0 % w/w. The reaction mass was filtered and the bed was washed with tetrahydrofuran (1 :2.5 w/v). The filtrate was quenched with 10 % acetic acid in water at 25±5°C. Reaction mass stirred for 50-60 minutes at 25±5°C. The filtrate was concentrated below 45°C until no distillate was observed. The mass was cooled to 5±5°C and stirred for 50-60 minutes. The reaction mass was filtered and the solid was taken in ethanol (1 :0.53 w/v). The reaction mass was cooled 0±5°C and stirred for 30-40 min. The solid was filtered and the bed was washed ethanol (1 :0.13 w/v). The solid was dried at 40±5 °C.

Step 3 – Synthesis of intermediate Br-IPM:

P

o

M
W = 286.7 MW = 20₄.9 Purified water (SLV, RGT)

Acetone (SLV)

IPC: NMT 1.0% water by KF

2-Bromoethylamine hydrobromide (1 : 1.0 w/w) and POBr^ (1 :0.7 w/w) were taken in DCM (1 :2 w/v) under nitrogen atmosphere. The reaction mixture was cooled to -70±5°C. Triethylamine (1 : 1.36 w/v) in DCM (1 :5 w/v) was added to the reaction mass at -70±5°C. The reaction mass was stirred for additional 30 min at -70±5°C. Reaction mass was warmed to 0±3°C and water (1 :1.72 w/v) was added. The reaction mixture was stirred at 0±3°C for 4 hrs. The solid obtained was filtered and filter cake was washed with ice cold water (2 x 1 :0.86 w/v) and then with chilled acetone (2 x 1 :0.86 w/v). The solid was dried in at 20±5°C.

Step 4 Synthesis ofTH-302

TH-302

MW = ₄₄9.0

Imidazole alcohol (IA) (1 : 1.0 w/w), Bromo-IPM (1 :2.26 w/w) and

triphenylphosphine (1 :2.0 w/w) were added to THF (1 : 13.5 w/v) at 25±5°C. The reaction

mass was cooled to 0±5°C and DIAD (1.5 w/v) was added. The reaction mixture warmed to 25±5°C and stirred for 2 hours. Progress of the reaction was monitored by HPLC. Solvent was removed below 50°C under vacuum. Solvent exchange with acetonitrile (1 :10.0 w/v) below 50°C was performed. The syrupy liquid was re-dissolved in acetonitrile (1 : 10.0 w/v) and the mixture was stirred at -20±5°C for 1 hour. The resulting solid was filtered and the filtrate bed was washed with chilled acetonitrile (1 : 1.0 w/v). The acetonitrile filtrate was concentrated below 50°C under vacuum. The concentrated mass was re-dissolved in ethyl acetate (1 : 10.0 w/v) and concentrated below 50°C under vacuum. The ethyl acetate strip off was repeated two more times. Ethyl acetate (1 : 10.0 w/v) and silica gel (230-400 mesh, 1 :5.3 w/w) were added to the concentrated reaction mass. The mixture was concentrated below 40°C under vacuum. n-Heptane (1 :5.0 w/v) was charged to the above mass and the mixture was evaporated below 40°C under vacuum. n-Heptane (1 :5.0 w/v) was again added to the above mass and the solid was filtered and the bed was washed with n-heptane (1 : 1.0 w/v). The solid was suspended in a mixture oftoluene (1 :7.1 w/v) and n-heptane (1 :21.3 w/v), stirred at 35±5°C for 15-20 minutes, filtered off and the bed was washed with n-heptane

(1 : 1.0 w/v). The solid was re-suspended in a mixture of toluene (1 : 10.6 w/v) and n-heptane (1 : 10.6 w/v), stirred at 35±5°C for 15-20 minutes, filtered off and the bed was washed with n-heptane (1 : 1.0 w/v). The solid was suspended in acetone (1 : 19.0 w/v), stirred at 35±5°C for 15-20 minutes, filtered off and the bed was washed with acetone (1 : 1.0 w/v). The acetone washes were repeated 3 more times. Filtrates from the above acetone washings were combined and concentrated below 40°C under vacuum. The residue dissolved in ethyl acetate (1 : 10.0 w/v) and concentrated below 40°C under vacuum. The ethyl acetate strip off was repeated one more time. The residue was re-dissolved in ethyl acetate (1 :5.5 w/v), cooled to 0±3°C and stirred at 0±3°C for 2 h and then at -20±5°C for 2 h. The solid was filtered and the solid was washed with ethyl acetate (1 :0.10 w/v). The solid was dissolved in ethyl acetate (1 : 10.0 w/v) at 50±5°C and the resulting solution was filtered through a cartridge filter. The filtrate was concentrated to ~4.0 w/w and stirred at 0±3°C for 4 hours. The solid was filtered and washed with ethyl acetate (1 :0.10 w/v). The crystallization from ethyl acetate was repeated and TH-302 was dried at 25±5°C. Table 2 shows how the process reduces solvent use.

Table 2: Solvent and Silica Gel Usage for 10 kg Column and 10 kg Column-free Purification

“Amounts are estimated from a 5 kg batch

^b Amounts are estimated

Example 2: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA was prepared according to the method described in Example 1. In a vessel, water (1 :7.0 w/v) was added. Dry crude ICA was added to the water. The reaction mixture was heated to 85±5°C until a clear solution was obtained. The reaction mass was cooled to 20±5°C and filtered through a Celite bed. The filter bed was washed with 2 x 5.0 of n-heptane. The material was dried for 2 hours at 25±5°C and then 45±5°C. As can be seen below, the number and levels of impurities decreased.

Table 3: Purity and Impurity Profile Comparison of Typical Crude ICA and Purified

ICA

Example 3: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA was prepared according to the method described in Example 1. In a vessel

ethanol (1 :30.0 w/v) and ICA (1 : 1.0 w/w) were mixed. The reaction mixture was stirred at

25±5°C for 30 minutes and filtered. Water (1 :50.0 w/v) was added and the mixture was

stirred at 50±5°C for 30 minutes. The reaction mass was cooled to 20±5°C and filtered. The isolated solid was dried at 25±5°C for 24 hours. As can be seen below, the number and levels

of impurities generally decreased.

Table 4: Purity and Impurity Profile Comparison of Typical Crude ICA and Purified

ICA

Example 4: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA was prepared according to the method described in Example 1. In a vessel

acetonitrile (1 :20.0 w/v) and ICA (1 : 1.0 w/w) were mixed at 25±5°C for one hour. The

reaction mixture was filtered and the solution was concentrated to ~ 6 volumes. The mixture

was then cooled to 0±5°C, stirred at this temperature for one hour and filtered. The isolated

solid was dried at 25±5°C for 24 hours. As can be seen below the number of impurities

decreased and except for TH-2717, the amounts also decreased.

Table 5: Purity and Impurity Profile Comparison of Typical Crude ICA and Purified

ICA

Example 5: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA is prepared according to the method described in Example 1 and purified by treatment with dimethylacetamide and water.

Example 6: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA is prepared according to the method described in Example 1 and purified by treatment with dimethylforamide and water.

Example 7: Synthesis ofTH-302 using alternative procedure to purify ICA:

[0109] Crude ICA is prepared according to the method described in Example 1 and purified by crystallization from a 1,4-dioxane and water mixture.

Example 8: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA is prepared according to the method described in Example 1 and purified by crystallization from a mixture of ethylene glycol and water.

Example 9: Synthesis ofTH-302 using alternative procedure to purify ICA:

Crude ICA is prepared according to the method described in Example 1 and purified by treatment with 2-propanol and water.

Example 10: Synthesis ofTH-302 using alternative procedure to purify ICA:

[0112] Crude ICA is prepared according to the method described in Example 1 and purified by treatment with 1-propanol and water.

Example 11: Synthesis ofTH-302 using alternative procedure to purify ICA:

[0113] Crude ICA is prepared according to the method described in Example 1 and purified by crystallization from a mixture of tetrahydrofuran and water.

Example 12: Synthesis ofTH-302 using alternative procedure to quench IA:

[0114] The reduction of ICA to IA was carried out according to Example 1 except that after reaction completion and filtration of the inorganics, the filtrate was quenched with 1.5 M hydrochloric acid.

Example 13: Synthesis ofTH-302 using alternative procedure to quench IA:

[0115] The reduction of ICA to IA was carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate was quenched with 1.5 M

hydrobromic acid.

Example 14: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA was carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate was quenched with

hydrobromic acid in acetic acid.

Example 15: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA was carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate was treated with sodium

hydrogen phosphate.

Example 16: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA was carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate was quenched with 10% acetic

acid in tetrahydrofuran.

Example 17: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA was carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate was quenched with water.

Example 18: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA is carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate is quenched with sulfuric acid.

Example 19: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA is carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate is quenched with citric acid.

Example 20: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA is carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate is treated with carbonic acid.

Example 21: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA is carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate is treated with phosphoric

acid.

Example 22: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA is carried out according to Example 1 except that after

reaction completion and filtration of the inorganics, the filtrate is quenched with oxalic acid.

Example 23: Synthesis ofTH-302 using alternative procedure to quench IA:

The reduction of ICA to IA is carried out according to Example 1 except that after reaction completion and filtration of the inorganics, the filtrate is quenched with boric acid.

Example 24: Synthesis ofTH-302 using alternative procedure to purify TH-302:

[0126] Coupling of bromo-IPM and IA was performed according to Example 1 except that after concentration of the reaction mixture, ethyl acetate (1 : 10 w/v) was added to the concentrated mass. The mixture was stirred at -55±5°C for 2 hours. The resulting solid was filtered and washed with chilled EtOAc (1 :2.0 w/v). The solid was reslurried in ethyl acetate (1 : 10 w/v) at -55±5°C for 2 hours, filtered and the solid was washed with chilled ethyl acetate (1 : 1.0 w/v). The filtrates from both filtrations were combined and treated with silica gel (1 :5.3 w/w) of silica gel (230-400 mesh). The mixture was concentrated below 40°C under vacuum. n-Heptane (1 :5.0 w/v) was again added to the above mass and the solid was filtered and the bed was washed with n-heptane (1 : 1.0 w/v). The solid was suspended in a mixture of toluene (1 :7.1 w/v) and n-heptane (1 :21.3 w/v), stirred at 35±5°C for 15-20 minutes, filtered off and the bed was washed with n-heptane (1 : 1.0 w/v). The solid was re-suspended in a mixture of toluene (1 : 10.6 w/v) and n-heptane (1 :10.6 w/v), stirred at 35±5°C for 15-20 minutes, filtered off and the bed was washed with n-heptane (1 : 1.0 w/v). The solid was suspended in acetone (1 : 19.0 w/v), stirred at 35±5°C for 15-20 minutes, filtered off and the bed was washed with acetone (1 : 1.0 w/v). The acetone washes were repeated 3 more times. Filtrates from the above acetone washings were combined and concentrated below 40°C under vacuum. The residue dissolved in ethyl acetate (1 :5.5 w/v), cooled to 0±3°C and stirred at 0±3°C for 2 h and then at -20±5°C for 2 h. The solid was filtered and the solid was washed with ethyl acetate (1 :0.10 w/v). The solid was dissolved in ethyl acetate (1 :27 w/v), stirred at 50±5°C and filtered through Celite. The filtrate was concentrated to ~4.0 w/w and stirred at 0±5°C for 4 hours. The recrystallization from ethyl acetate was repeated and TH- 302 was dried at 25±5°C. Table 4 shows how the process reduced solvent use.

Table 4: Estimated Solvent and Silica Gel Usage for Column and 10 kg Column-free

(EtOAc) Purification

References

Evofosfamide

Names
IUPAC name (1-Methyl-2-nitro-1H-imidazol-5-yl)methyl N,N’-bis(2-bromoethyl)phosphorodiamidate
Other names TH-302; HAP-302
Identifiers
CAS Registry Number	918633-87-1
ChemSpider	10157061
InChI[show]
Jmol-3D images	Image
PubChem	11984561
SMILES[show]
Properties
Molecular formula	C9H16Br2N5O4P
Molar mass	449.04 g·mol−1
Solubility in water	6 to 7 g/l

///////////Orphan Drug Status, soft tissue sarcoma,  Pancreatic cancer, Fast track,  TH-302, TH 302, эвофосфамид ,  إيفوفوسفاميد ,  艾伏磷酰胺 , Evofosfamide, 918633-87-1, PHASE 3

O=[N+]([O-])c1ncc(COP(=O)(NCCBr)NCCBr)n1C

Continuous Flow Stereoselective Synthesis of (S)-Warfarin

The same catalytic packed-bed reactor was used for the preparation of (S)-warfarin 107 under continuous flow conditions (Scheme ).A solution of 4-OH-coumarin 104, benzalacetone105, and trifluoroacetic acid as a cocatalyst in dioxane was flowed into the reactor containing the polystyrene-supported 9-amino-epi-quinine 122. With a residence time of 5 h at 50 °C, we were able to isolate the product in up to 90% yield and up to 87% ee. Further studies are needed in order to optimize the reaction under continuous flow conditions; however, the proposed protocol already offers the possibility to extend catalyst’s lifetime, longer than in batch mode, further suggesting interesting future applications for the catalytic reactors.

The Pericàs group published the stereoselective Michael addition of ethyl nitroacetate to benzalacetone promoted by polystyrene-supported 9-amino-9-deoxy-epi-quinine 126 under continuous flow conditions. It should be pointed out that the polystyrene in our hands is a highly reticulated, insoluble polymer, while the polystyrene used by the Pericàs group is a swelling resin; a careful choice of the reaction solvent should be done, as this may affect the reaction course. The functionalized resin was packed into a Teflon tube between two plugs of glass wool. The reaction was run by pumping a solution of the two reagents and benzoic acid as a cocatalyst in CHCl₃ (chosen after careful solvent screening) at 30 °C for 40 min residence time. Notably, 3.6 g (12.9 mmol) of the desired adducts were collected in 21 h of operation in roughly 1/1 dr and 97/98% ee.

Porta, R.; Benaglia, M.; Puglisi, A. Unpublished results.

Izquierdo, J.; Ayats, C.; Henseler, A. H.; Pericàs, M. A. Org. Biomol. Chem. 2015, 13, 4204, DOI: 10.1039/C5OB00325C

A polystyrene (PS)-supported 9-amino(9-deoxy)epi quinine derivative catalyzes Michael reactions affording excellent levels of conversion and enantioselectivity using different nucleophiles and structurally diverse enones. The highly recyclable, immobilized catalyst has been used to implement a single-pass, continuous flow process (residence time: 40 min) that can be operated for 21 hours without significant decrease in conversion and with improved enantioselectivity with respect to batch operation. The flow process has also been used for the sequential preparation of a small library of enantioenriched Michael adducts.

NMR 

General Data:

Chemical Names:	4-hydroxy-3-(3-oxo-1-phenyl-butyl)-chromen-2-one 3-(2-acetyl-1-phenylethyl)-4-hydroxycoumarin (+ -)Warfarin
Formula:	C19H16O4
CAS Number:	81-81-2
Molecular Weight:	308.33
Structure:
Isomers:	Optical Isomers: S-Warfarin and R-Warfarin
Melting Point /ºC :	161
Optical Rotation:	S-Warfarin : -25.5 ± 1º R-Warfarin : +24.8 ± 1º

 

Tautomerization of warfarin substructures, whose combination generates 40 distinct tautomeric forms of warfarin

The stereochemical assignment of (−)-(S)-warfarin was initially achieved by relating it to (−)-(R)-beta-phenylcaproic acid through a series of reactions not involving the asymmetric center {B.D.West, S.Preis, C.H.Schroder, & K.P.Link, J.Amer.Chem.Soc.,1961,83, 2676}. This assignment was confirmed by a determination of the crystal and molecular structure, and using the anomalous scattering of oxygen, and absolute configuration of (−)-(S)-Warfarin was measured {E.J.Valente, W.F.Trager and L.H.Jensen, Acta Cryst. 1975. B31, 954}.

The Hemiketal

The primary feature of the structure of (−)-warfarin is the hemiketal ring formed by cyclization of the side-chain keto function and the phenolic hydroxyl in the 4 position of the coumarin ring system. The crystal structure of racemic warfarin has the same feature. In solution n.m.r. spectra shows that the hemiketal is present in acetone solution.

Bond Lengths

The hemiketal bonding is rather weak. Thus the bond lengths within the hemiketal show that the atoms retain some of the characteristic of an open side chain keto group.

The Absolute Configuration

In the open chain keto form warfarin has two isomers, R andS, however the hemiketal introduces a second assymmetric center, so that we can have RR,SS, RS, and SR forms. The crystal structure determination favoured the SS enantiomer in the crystal studied.

Enantiomers & Biochemical Function

The S-isomer is very much more potent than the R isomer in both rats and humans.The S-isomer is stereoselectively oxidized to the inactive 7-hydroxywarfarin, and the keto-group of the R-isomer is stereospecifically reduced to the slightly active R,S-alcohol. Both isomers are oxidized to the inactive 6-hydroxywarfarin.

It is evident that we are dealing with a very complex system indeed; the presence of the hemiketal adds four more enantiomers to the complexity pot. Recent work has unravelled some more of the mechanisms behind the Vitamin K1 antagonism of Warfarin.

//////////////////////Continuous Flow,  Stereoselective Synthesis, (S)-Warfarin, FLOW CHEMISTRY, FLOW SYNTHESIS

SNS 032, BMS-387032

N-[5-[[[5-(1,1-Dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4-piperidinecarboxamide

Cas 345627-80-7, MP 165-167° C

M.Wt:380.53, Formula:C₁₇H₂₄N₄O₂S₂

SNS 032, BMS-387032 HYDROCHLORIDE

A potent and selective Cdk inhibitor

Potent inhibitor of cyclin-dependent kinases (cdks) 9, 2 and 7 (IC₅₀ values are 4, 38 and 62 nM respectively). Displays no activity against 190 additional kinases (IC₅₀ >1000 nM). Arrests the cell cycle at G₂/M; inhibits transcription, proliferation and colony formation, and induces apoptosis in RPMI-8226 multiple myeloma cells. Prevents tumor cell-induced VEGF secretion and in vitro angiogenesis. SNS-032 (BMS-387032) has firstly been described as a selective inhibitor of CDK2 with IC50 of 48 nM in cell-free assays and is 10- and 20-fold selective over CDK1/CDK4. It is also found to be sensitive to CDK7/9 with IC50 of 62 nM/4 nM, with little effect on CDK6. Phase 1.

Quality Control & MSDS

COA NMR HPLC Datasheet SDS/MSDS

• COA (Certificate Of Analysis)
• HPLC
• NMR (Nuclear Magnetic Resonance)
• MSDS (Material Safety Data Sheet)

SNS-032 (BMS-387032) is a potent and selective inhibitor of cyclin-dependent kinases (CDKs) 2, 7, and 9 [1], with IC50 values of 38 nM, 62 nM and 4 nM, respectively [2].

CDKs mean a family of serine/threonine kinases regulating cell cycle process. Some CDKs are related to transcription control and are often perturbed in cancer cells [3].

Decrease in the phosphorylation at Ser5 and Ser2 in the C-terminal domain (CTD) of RNA Pol II can indicate the inhibition to CDK9 and CDK7 [1]. Chronic lymphocytic leukemia (CLL) cells treated with SNS-032 for 6 or 24 hours showed a decrease in the phosphorylation of Ser2 and Ser5 of the CTD of RNA Pol II, this appeared to be both time- and concentration- dependent, and remarkably consistent among samples. For the phosphorylation of Ser2, the inhibition of SNS-032 was greater than that for the phosphorylation of Ser5, this was consistent with the fact that IC50 for the inhibition of CDK9 was lower compared with that for the inhibition of CDK7 (4 nM vs 62 nM). After 6 hours of SNS-032 exposure, protein levels of CDK7 and CDK9 were stable, but declined at 24 hours [4].

In patients with chronic lymphocytic leukemia (CLL), infusion of SNS-032 in a total dose of 75 mg/m2 resulted in a decrease in the phosphorylation at Ser5 and Ser2 in the C-terminal domain of RNA Pol II. This indicated the inhibition to Cdk9 and Cdk7 by SNS-032. This inhibition was first seen 2 hours after the beginning of the infusion with SNS-032, was pronounced after 6 hours and returned to baseline after 24 hours [1].

SNS-032 (formerly BMS-387032) is a small-molecule cyclin-dependent kinase (CDK) inhibitor currently in phase I clinical trials for the treatment of B-cell malignancies and advanced solid tumors. Preclinical studies have shown that SNS-032 is a specific and potent inhibitor of CDK2, 7 and 9 which induces cell cycle arrest and apoptosis in tumor cell lines. It was shown to inhibit in vitro angiogenesis and prostaglandin E2 (PGE2) production, both strongly associated with tumorigenesis. Phase I clinical trials support the safety and tolerability of SNS-032 as evaluated in dose-escalation studies. The compound is currently administered by i.v. infusion but has shown promising potential for oral delivery.

NMR

CLIP

The structures of representative protein kinases inhibitors based on the aminopyrazole scaffold.http://www.mdpi.com/1422-0067/14/11/21805/htm

CLIP

N-(Cycloalkylamino)acyl-2-aminothiazole Inhibitors of Cyclin-Dependent Kinase 2. N-[5-[[[5-(1,1-Dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4- piperidinecarboxamide (BMS-387032), a Highly Efficacious and Selective Antitumor Agent, 

N-Acyl-2-aminothiazoles with nonaromatic acyl side chains containing a basic amine were found to be potent, selective inhibitors of CDK2/cycE which exhibit antitumor activity in mice. In particular, compound 21 {N-[5-[[[5-(1,1-dimethylethyl)-2-oxazolyl]methyl]thio]-2-thiazolyl]-4-piperidinecarboxamide, BMS-387032}, has been identified as an ATP-competitive and CDK2-selective inhibitor which has been selected to enter Phase 1 human clinical trials as an antitumor agent. In a cell-free enzyme assay, 21 showed a CDK2/cycE IC₅₀ = 48 nM and was 10- and 20-fold selective over CDK1/cycB and CDK4/cycD, respectively. It was also highly selective over a panel of 12 unrelated kinases. Antiproliferative activity was established in an A2780 cellular cytotoxicity assay in which 21 showed an IC₅₀ = 95 nM. Metabolism and pharmacokinetic studies showed that 21 exhibited a plasma half-life of 5−7 h in three species and moderately low protein binding in both mouse (69%) and human (63%) serum. Dosed orally to mouse, rat, and dog, 21showed 100%, 31%, and 28% bioavailability, respectively. As an antitumor agent in mice, 21administered at its maximum-tolerated dose exhibited a clearly superior efficacy profile when compared to flavopiridol in both an ip/ip P388 murine tumor model and in a sc/ip A2780 human ovarian carcinoma xenograft model.

CLIP

http://pubs.rsc.org/en/content/articlehtml/2016/md/c6md90040b

Heat shock factor 1 (HSF1) is a transcription factor that plays key roles in cancer, including providing a mechanism for cell survival under proteotoxic stress. Therefore, inhibition of the HSF1-stress pathway represents an exciting new opportunity in cancer treatment. We employed an unbiased phenotypic screen to discover inhibitors of the HSF1-stress pathway. Using this approach we identified an initial hit (1) based on a 4,6-pyrimidine scaffold (2.00 μM). Optimisation of cellular SAR led to an inhibitor with improved potency (25, 15 nM) in the HSF1 phenotypic assay. The 4,6-pyrimidine 25 was also shown to have high potency against the CDK9 enzyme (3 nM).

COMPD 25

1H NMR (500 MHz, DMSO-d6) δ 10.38 (s, 1H), 9.21 (s, 1H), 8.74 (d, J = 0.9 Hz, 1H), 8.62 (dd, J = 8.2, 1.5 Hz, 1H), 8.56 (dd, J = 4.7, 1.5 Hz, 1H), 8.16-8.13 (m, 2H), 7.64 (br d, J = 8.6 Hz, 1H), 7.52-7.47 (m, 2H), 4.14 (t, J = 5.9 Hz, 2H), 2.66 (t, J = 5.9 Hz, 2H), 2.47-2.42 (m, 4H), 1.53-1.47 (m, 4H), 1.42 – 1.33 (m, 2H). 13C NMR (126 MHz, DMSO-d6) δ 160.74, 158.32, 156.72, 154.88, 150.74, 146.47, 145.38, 143.74, 134.21, 125.02, 124.16, 122.29, 119.60, 114.32, 94.06, 66.49, 57.35, 54.35, 25.54, 23.88. HRMS (ESI+ ): calcd for C22H25N8O (M + H)+ , 417.2146; found 417.2163.

NOTE, THERE IS ERROR IN STRUCTURE ABOVE OF SNS 032

CC(C)(C)C1=CN=C(O1)CSC2=CN=C(S2)NC(=O)C3CCNCC3

SR48692 (Meclinertant)

Reminertant; SR 48692

CAS [146362-70-1]

• Molecular FormulaC₃₂H₃₁ClN₄O₅
• Average mass587.065

SEE…...https://newdrugapprovals.org/2014/12/31/meclinertant-sr48692/

2-[[1-(7-chloroquinolin-4-yl)-5-(2,6-dimethoxyphenyl)pyrazole-3-carbonyl]amino]adamantane-2-carboxylic acid

• Originatorsanofi-aventis
• ClassAnalgesics; Antineoplastics; Antipsychotics
• Mechanism of ActionNeurotensin antagonists

Meclinertant (SR-48692) is a drug which acts as a selective, non-peptide antagonist at the neurotensin receptor NTS₁, and was the first non-peptide antagonist developed for this receptor.^[1][2] It is used in scientific research to explore the interaction between neurotensin and other neurotransmitters in the brain,^{[3][4][5][6][7][8]} and produces anxiolytic, anti-addictive and memory-impairing effects in animal studies.^{[9][10][11][12]}

CLIP

Methods for the synthesis of pharmaceuticals have improved over the years, however, the technology and tools used to perform synthetic operations have remained the same. Batch-mode processes are still common but many improvements can be made by using modern technologies. Recently, the use of machine-assisted protocols has increased, with flow-based chemical synthesis being extensively investigated. Under dynamic flow regimes, mixing and heat transfer can be more accurately controlled, the use of solid-phase reagents and catalysts can facilitate purification, and tedious downstream processes (workup, extraction, and purification) are reduced.
Steven V. Ley and co-workers, University of Cambridge, UK, have been evaluating the utility of flow-based syntheses to accelerate multistep routes to highly complex, medically relevant compounds, in this case Meclinertant (SR48692, pictured). They show that new technologies can help to overcome many synthetic issues of the existing batch process. In this case, flow chemistry has allowed control of exothermic events, controlled the superheating of solvents, and streamlined the synthesis by allowing reaction telescoping. It has also helped to prevent back mixing and the accumulation of byproducts. The use of polymer-supported reagents has simplified downstream processing and enhanced the safety of reactions, and in-line monitoring can track hazardous intermediates.

These new technologies have been shown to be powerful synthetic tools, although care must be taken not to convert them to expensive solutions to nonexistent problems.

http://community.dur.ac.uk/i.r.baxendale/papers/ChemEurJ2013.19.7917.pdf

A Machine-Assisted Flow Synthesis of SR48692: A Probe for the Investigation of Neurotensin Receptor-1,
Claudio Battilocchio, Benjamin J. Deadman, Nikzad Nikbin, Matthew O. Kitching, Ian R. Baxendale, Steven V. Ley,
Chem. Eur. J. 2013.
DOI: 10.1002/chem.201300696

2-[1-(7-Chloroquinolin-4-yl)-5-(2,6-dimethoxyphenyl)-1H-pyrazole-3-carboxamido]adamantane-2-carboxylic acid (1):

Polymer-supported sulfonic acid (QP-SA; 0.6 g, 2.4 mmol) was added to a solution of tert-butyl 2-[1- (7-chloroquinolin-4-yl)-5-(2,6-dimethoxyphenyl)-1H-pyrazole-3-carboxamido]adamantane-2-carboxylate (13; 30 mg, 0.05 mmol) in dichloromethane and the reaction was stirred at RT for 18 h. The QP-SA was filtered off and the filtrate concentrated in vacuo to provide the title compound as white crystals (yield 25 mg, 0.04 mmol, 86%).

M.p. 219–222 deg C;

1 H NMR (400 MHz, CDCl3, 25 deg C): d=8.91 (d, 1H, J=4.6 Hz), 8.15 (d, 1H, J=2.1 Hz), 7.78 (d, 1H, J=9.1 Hz), 7.68 (dd, 1H, J=2.1, 9.1 Hz), 7.28 (d, 1H, J=4.7 Hz), 7.24 (t, 1H, J=8.5 Hz), 7.91 (s, 1H), 6.52 (d, 2H, J=8.5 Hz), 3.42 (s, 6H), 2.64–2.56 (m, 2H), 2.17–2.05 (m, 2H), 2.04–1.92 (m, 2H), 1.82–1.71 (m, 2H), 1.71–1.61 (m, 4H), 1.61–1.50 ppm (m, 2H); 

13C NMR (100 MHz, CDCl3, 25 deg C): d=173.3(C), 159.9 (C), 157.5 (C), 157.5 (C), 151.8 (CH), 149.1 (C), 143.4 (C), 139.2 (C), 134.8 (C), 131.9 (CH), 128.0 (CH), 127.7 (CH), 125.9 (CH), 122.2 (C), 118.6 (CH), 109.6 (CH), 105.8 (C), 104.0 (CH), 55.4 (CH3), 55.3 (C), 37.4 (CH2), 33.6 (CH2), 32.8 (CH2), 31.9 (CH), 26.5 (CH), 26.2 ppm (CH);

FT-IR (neat): 3405, 2922, 1728, 1674, 1591, 1527, 1474, 1433, 1379, 1357, 1288, 1251, 1206, 1101, 1077, 1031, 1006, 957, 882, 865, 823, 779, 725, 682 cm1 ;

LCMS: tR =5.29 min, m/z [M+H]+: 587.46;

HRMS (ESI): m/z calcd for C32H32N4O5Cl+: 587.2061, found 587.2053; the structure was unambiguously confirmed by single X-ray crystallography; space group P1¯: a= 10.249, b=11.718, c=12.634 ; a=76.6, b=72.9, g=76.4o

CLIP AND ITS OWN REFERENCES

Although batch processes remain the most used procedure for running chemical reactions, the use of machine-assisted flow methodologies(24) enables an improved efficiency and high throughput. A direct comparison between conventional batch preparation and flow multistep synthesis of selective neurotensine probe SR48692 (Meclinertant) was reported by Ley and co-workers in 2013 (Scheme 6).(25)

In this case study, the authors investigated whether flow technology could accelerate a multistep synthesis (i.e., higher yields or lower reaction times) and overcome many synthetic issues (i.e., solid precipitation or accumulation of byproducts). The initial Claisen condensation between ketone 31 and ethyl glyoxalate in the presence of NaOEt as base and EtOH as solvent in batch is run at room temperature and product 32 is obtained in 60% yield after 3 h stirring.

Superheating (heat above solvent boiling point) the reaction in flow provided a faster alternative: using a 52 mL PFA reactor coil at 115 °C with a residence time of 22 min gave the corresponding product 32 in 74% yield. In order to solve some problems of solid accumulation an ad-hoc pressurized stainless-steel tank (5 bar, nitrogen) was designed; it allowed to run the reaction continuously without any precipitation or blockage.

The following reaction between 32 and commercially available hydrazine 33 was performed in DMF in the presence of concentrated H₂SO₄. After 52 min of residence time at 140 °C into a 52 mL PFA reactor coil the crude mixture was treated with an Na₂CO₃ aq. and then inline extracted through a semipermeable membrane with CH₂Cl₂. After crystallization, pyrazole ester 34 was isolated in 89% yield.

The corresponding reaction in batch was conducted in DMF under microwaves irradiation at 140 °C for 2 h. Running the reaction in batch on the same scale as in flow (3.58 mmol) gave product 34 in a lower yield (70%). The subsequent hydrolysis was performed combining a THF solution of ester 34 and 3 M aqueous KOH. The reaction was performed inside a 14 mL PFA reactor coil heated at 140 °C with a residence time of 14 min.

Upon treatment with 3 M HCl aq., acid 35 precipitated, and it was isolated by filtration in 90% yield. In this case, the corresponding batch hydrolysis afforded product 35 with the same yield (90%); however, a longer reaction time (1.5 h) was required. The final amide formation was performed by reacting acid 35 (activated as acyl chloride) and protected amino alcohol 37through a telescoped synthesis. Triphosgene 36 (a safer substitute for phosgene) was found to be the best acid activator.

Triphosgene decomposition occurred in the presence of DIPEA at 100 °C into a stainless steel heat exchanger, where phosgene was generated. The crude mixture, containing also acid 35, then passed into a 2.5 mL stainless steel reactor coil at 25 °C, to complete the formation of the corresponding acyl chloride. An inline Flow-IR spectrometer(26)was used to monitor the formation of phosgene without exposing the operator to the toxic gas during analysis. As soon as acyl chloride was formed it was reacted with protected amino alcohol 37.

The amide formation took place into a 14 mL stainless steel reactor coil at 100 °C with a residence time of 75 s. Amide 38 was isolated in 85% yield after quenching with NH₄Cl and extraction with AcOEt. For obvious safety concerns, avoiding the handling of phosgene and the isolation of highly reactive acyl chloride intermediate represent a remarkable improvement with respect to batch procedure.

Finally, meclinertant 39 was obtained after deprotection of ester38 by using a polymer-supported sulfonic acid. The last synthetic step was conducted in batch on a small scale; however, it could be easily transferred to flow mode by using a column packed with commercially available polymer-supported sulfonic acid.

24 Ley, S. V.; Fitzpatrick, D. E.; Myers, R. M.; Battilocchio, C.; Ingham, R. J. Angew. Chem., Int. Ed. 2015, 54, 2, DOI: 10.1002/anie.201501618

25.Battilocchio, C.; Deadman, B. J.; Nikbin, N.; Kitching, M. O.; Baxendale, I. C.; Ley, S. V. Chem. – Eur. J. 2013, 19, 7917, DOI: 10.1002/chem.201300696

CLIP AND ITS OWN REFERENCES

The choice of the flow reactor also plays a key role in the synthesis of meclinertant (SR48692, 103), which is a potent probe for investigating neurotensin receptor-1 [92]. The flow synthesis of this challenging compound was reported in 2013 and aims to evaluate the benefits of flow chemistry in order to avoid shortcomings of previous batch synthesis efforts particularly in regard to scale up [93].

The investigation first involved the preparation of the key acetophenone starting material 112 which although commercially available was expensive and could be generated from 1,3-cyclohexadione (104). The sequence consisted of O-acetylation, a Steglich rearrangement, oxidation and a final methylation reaction.

As the use of flow chemistry had already improved the O-acetylation during scale-up tests (130 mmol) by avoiding exotherms, it was anticipated that the subsequent Steglich rearrangement could be accomplished in flow using catalytic DMAP instead of stoichiometric AlCl₃ as precedented (Scheme 19).

This was eventually realised by preparing a monolithic flow reactor functionalised with DMAP that proved far superior to commercially available DMAP on resin. Employing the monolithic reactor cleanly catalysed the rearrangement step when a solution of 106 was passed through the reactor at elevated temperature (100 °C, 20 min residence time).

The resulting triketone 107 was telescoped into an iodine mediated aromatisation, followed by high temperature mono-methylation using dimethyl carbonate/dimethylimidazole as a more benign alternative to methyl iodide at scale.

The subsequent Claisen condensation step between ketone 112 and diethyl oxalate (113) was reportedly hampered by product precipitation and clogging problems, thus a pressure chamber was developed [94] that would act as a pressure regulator allowing this step to be scaled up in flow in order to provide 114 on multigram scale (134 g/h).

A Knorr pyrazole formation between 114 and commercially available hydrazine 115 had previously been found difficult to scale up in batch (the yield dropped from 87% to 70%) and was thus translated into a high temperature flow protocol (140 °C) delivering the desired product 116 in 89% yield (Scheme 20).

Ester hydrolysis and a triphosgene (118) mediated amide bond formation between acid 117 and adamantane-derived aminoester119 [95] completed this flow synthesis. Meclinertant (103) was subsequently obtained after batch deprotection using polymer supported sulfonic acid.

Overall, this study showcases how flow chemistry can be applied to gain benefits when faced with problems during mesoscale synthesis of a complex molecule. However, despite the successful completion of this campaign, it could be argued that the development time required for such a complex molecule in flow can be protracted; therefore both synthetic route and available enabling technologies should be carefully examined before embarking upon such an endeavour.

Marcus Baumann and Ian R. Baxendale

Department of Chemistry, Durham University, South Road, DH1 3LE Durham, United Kingdom

Corresponding author email

Associate Editor: J. A. Murphy

Beilstein J. Org. Chem.2015,11, 1194–1219.

EP 0477049; FR 2665898; JP 1992244065; US 5420141; US 5607958; US 5616592; US 5635526; US 5744491; US 5744493

The condensation of 2′,6′-dimethoxyacetophenone (I) with diethyl oxalate (II) by means of sodium methoxide in refluxing methanol gives the dioxobutyrate (III), which is cyclized with 7-chloroquinoline-4-hydrazine (IV) in refluxing acetic acid yielding the pyrazole derivative (V). The hydrolysis of the ester group of (V) with KOH in refluxing methanol/water affords the corresponding carboxylic acid (VI), which is finally treated with SOCl2 in refluxing toluene and condensed with 2-aminoadamantane-2-carboxylic acid.

Meclinertant

Systematic (IUPAC) name
2-([1-(7-Chloro-4-quinolinyl)-5-(2,6-dimethoxyphenyl)-1H-pyrazole-3-carbonyl]amino)admantane-2-carboxylic acid
Identifiers
CAS Number	146362-70-1
PubChem	CID 119192
IUPHAR/BPS	1582
UNII	5JBP4SI96H
ChEMBL	CHEMBL506981
Chemical data
Formula	C32H31ClN4O5
Molar mass	587.064

References

//////////////////////Flow synthesis, Meclinertant, SR48692, Reminertant,  SR 48692, 146362-70-1

COC1=C(C(=CC=C1)OC)C2=CC(=NN2C3=C4C=CC(=CC4=NC=C3)Cl)C(=O)NC5(C6CC7CC(C6)CC5C7)C(=O)O

Almorexant (INN, codenamed ACT-078573) is an orexin antagonist, functioning as a competitive receptor antagonist of the OX₁ and OX₂ orexin receptors, which was being developed by the pharmaceutical companies Actelion and GSK for the treatment of insomnia. Development of the drug was abandoned in January 2011.^[1]

Development

Originally developed by Actelion, from 2007 almorexant was being reported as a potential blockbuster drug, as its novel mechanism of action (orexin receptor antagonism) was thought to produce better quality sleep and fewer side effects than the traditionalbenzodiazepine and z drugs which dominated the multibillion-dollar insomnia medication market.^[2][3]

In 2008, pharmaceutical giant GlaxoSmithKline bought the development and marketing rights for almorexant from Actelion for an initial payment of $147 million.^[4] The deal was worth a potential $3.2billion if the drug were to successfully complete clinical development and obtain FDA approval.^[5] GSK and Actelion continued to develop the drug together, and completed a Phase IIIclinical trial in November 2009.^[6]

However, in January 2011 Actelion and GSK announced they were abandoning the development of almorexant because of its side effect profile.^[1][7]

Mechanism of action

Almorexant is a competitive, dual OX₁ and OX₂ receptor antagonist and selectively inhibits the functional consequences of OX₁ and OX₂ receptor activation, such as intracellular Ca²⁺ mobilization.

PAPER

http://pubs.rsc.org/en/content/articlelanding/2013/ob/c3ob40655e#!divAbstract

An enantioselective synthesis of almorexant, a potent antagonist of human orexin receptors, is presented. The chiral tetrahydroisoquinoline core structure was prepared via iridium-catalysed asymmetric intramolecular allylic amidation. Further key catalytic steps of the synthesis include an oxidative Heck reaction at room temperature and a hydrazine-mediated organocatalysedreduction.

PATENT

https://www.google.com/patents/EP2227454A2?cl=en

Reaction scheme 5:

7*CH₃COOH

Step 11 : synthesis of (2R)-2-{(-/S)-6,7-dimethoxy-1 -[2-(4-thfluoromethyl-phenyl)- ethyl]-3,4-dihydro-1 /-/-isoquinolin-2-yl}-Λ/-methyl-2-phenyl-acetamide (compound 8)

To the solution of the compound 7 in MIBK are added 1.2 equivalents of the compound 6, 1.1 equivalents caustic soda and 1.1 equivalents potassium carbonate and heated to 70-90 ⁰C. After full conversion the solution is cooled to RT and water is added. Phase separation is followed by a second washing of the organic phase with water and again phase separation. Step 12: synthesis of (2R)-2-{(-/S)-6,7-dimethoxy-1 -[2-(4-trifluoromethyl-phenyl)- ethyl]-3,4-dihydro-1 /-/-isoquinolin-2-yl}-/\/-nnethyl-2-phenyl-acetannide hydrochloride acid (compound I)

To the organic phase of step 11 is added 1 equivalent aqueous hydrochloric acid and then the water removed by azeotropic distillation in vacuo. The precipitate is dissolved by addition of 2-propanol at 75 ⁰C. Concentration of the solution leads to crystallisation and the suspension is then cooled to RT. To ensure complete crystallisation, the suspension is aged at RT, then filtered and washed with a MIBK-2-propanol mixture. The product is dried in vacuo at 50 ⁰C.

PAPER

Several methods are presented for the enantioselective synthesis of the tetrahydroisoquinoline core of almorexant (ACT-078573A), a dual orexin receptor antagonist. Initial clinical supplies were secured by the Noyori Ru-catalyzed asymmetric transfer hydrogenation (Ru-Noyori ATH) of the dihydroisoquinoline precursor. Both the yield and enantioselectivity eroded upon scale-up. A broad screening exercise identified TaniaPhos as ligand for the iridium-catalyzed asymmetric hydrogenation with a dedicated catalyst pretreatment protocol, culminating in the manufacture of more than 6 t of the acetate salt of the tetrahydroisoquinoline. The major cost contributor was TaniaPhos. By switching the dihydroisoquinoline substrate of the Ru-Noyori ATH to its methanesulfonate salt, the ATH was later successfully reduced to practice, delivering several hundreds of kilograms of the tetrahydroisoquinoline, thereby reducing the catalyst cost contribution significantly. The two methods are compared with regard to green and efficiency metrics.

Catalytic Asymmetric Reduction of a 3,4-Dihydroisoquinoline for the Large-Scale Production of Almorexant: Hydrogenation or Transfer Hydrogenation?

References

External links

• Actelion’s official website

Almorexant

Systematic (IUPAC) name
(2R)-2-[(1S)- 6,7-dimethoxy- 1-{2-[4-(trifluoromethyl)phenyl]ethyl}- 3,4-dihydroisoquinolin-2(1H)-yl]- N-methyl- 2-phenylacetamide
Clinical data
Routes of administration	Oral
Pharmacokinetic data
Metabolism	Hepatic
Identifiers
CAS Number	871224-64-5
ATC code	none
PubChem	CID 23727689
IUPHAR/BPS	2886
ChemSpider	21377865
UNII	9KCW39P2EI
ChEMBL	CHEMBL455136
Chemical data
Formula	C29H31F3N2O3
Molar mass	512.6 g/mol (free base)

///////Almorexant,  ACT-078573

CNC(=O)C(C1=CC=CC=C1)N2CCC3=CC(=C(C=C3C2CCC4=CC=C(C=C4)C(F)(F)F)OC)OC

1, 2-Bis(4-(4-4-nitrophenyl)piperazin-1-yl)ethanone

CAS 330633-91-5

CDRI-?

For treatment of androgen sensitive prostatic disorders

1, 2-Bis(4-(4-4-nitrophenyl)piperazin-1-yl)ethanone

In the quest for novel scaffolds for the management of androgen sensitive prostatic disorders like prostate cancer and benign prostatic hyperplasia, a series of twenty-six aryl/heteroaryl piperazine derivatives have been described. Three compounds, 8a, 8c and 9a, exhibited good activity profiles against an androgen sensitive prostate cancer cell line (LNCaP) with EC₅₀values of 9.8, 7.6 and 11.2 μM, respectively. These compounds caused a decrease in luciferase activity and a decline in PSA and Ca²⁺ levels, which are indicative of their anti-androgenic and α_1A-adrenergic receptor blocking activities, respectively.

Compound 9a reduced the prostate weight of rats (47%) and in pharmacokinetic analysis at 10 mg kg⁻¹ it demonstrated an MRT of ∼14 h post dose, exhibiting high levels in prostate. Compound 9a docked in a similar orientation to hydroxyflutamide on an androgen receptor and showed strong π–π interactions. These findings reveal that compound 9a is a promising candidate for management of prostatic disorders with anti-androgenic and α_1A-blocking activities.

1, 2-Bis(4-(4-4-nitrophenyl)piperazin-1-yl)ethanone (9a) To the mixture of 8a (0.3 g, 1.06 mmol) and Et3N (0.3 mL, 2.12 mmol) in CHCl3 (5 mL) was added 1-(4-nitrophenyl)piperazine (7a, 0.320 g, 1.59 mmol) in 5 mL CHCl3 dropwise within 1 h. After complete addition reaction mixture was further stirred in an oil bath at 80-85 °C for 15 h. The reaction mixture was cooled, washed with water (5 mL × 3) and the organic layer was separated. Combined organic layer was dried (anhyd. Na2SO4 and concentrated under reduced pressure in rotavapor. The solid obtained was purified by recrystallization using EtOAc/Hexane which furnished yellow crystals (yield 81%);

mp: 156-157 °C; IR (KBr)  (cm-1): 3019, 2399, 1640, 1597, 1506, 1423, 1330;

1H NMR (400 MHz, CDCl3):  8.14-8.09 (4H, m), 6.84-6.81 (4H, m), 3.84-3.83 (4H, m), 3.49-3.44 (8H, m), 3.33 (2H, s), 2.72 (4H, t, J = 5.0 Hz);

13C NMR (75.4 MHz, CDCl3):  167.7, 154.7, 154.3, 138.8, 138.4, 125.9, 125.8, 112.9, 112.7, 60.8, 52.5, 46.9, 46.7, 44.6;

HRMS (ESI positive) m/z calcd. for C22H26N6O5 [M+H]+ : 455.2043, found: 455.2034;

Anal calcd. for C22H26N6O5: C, 58.14; H, 5.77; N, 18.49, found: C, 58.31; H, 5.92; N, 18.66.

SONAL GUPTA

Medicinal & Process Chemistry Division, CSIR-Central Drug Research Institute, Sector 10, Jankipuram ext., Lucknow-226031, India

Dr. VISHNU LAL SHARMA

http://www.cdriindia.org/VL_Sharma.htm

Dr. VISHNU LAL SHARMA
	Senior Principal Scientist (CSIR-CDRI ) / Professor (AcSIR) Lab No. CSS-SF-201, Medicinal and Process Chemistry Division Central Drug Research Institute, B.S. 10/1, Sector 10, Jankipuram Extension, Sitapur Road Lucknow- 226031
Educational Qualifications M.Sc (Organic Chemistry, Lucknow University, Lucknow, Uttar Pradesh, 1978) Ph.D. (Chemistry, Lucknow University, Lucknow, Uttar Pradesh, 1985) Date of Birth February 7th, 1958 E- Mail vl_sharma@cdri.res.in, vlscdri@gmail.com Phone No. +91-0522-2772450/550, Ext. 4671. Mobile No. +91-9415074195 Fax No. +91-522-2771941 Research Experience (Area) Medicinal chemistry, Organic chemistry. Google Scholar https://scholar.google.co.in/citations?user=cAsQaiYAAAAJ&hl=en Research gate https://www.researchgate.net/profile/Vishnu_Sharma13
POST-DOCTORAL RESEARCH (ABROAD)
• University of Dusseldorf, Dusseldorf, Germany, Oct., 1994 to Dec., 1994
CURRENT AREAS OF INTEREST
• Medicinal Chemistry, Synthetic organic chemistry and Process chemistry. • The research focused in my group is related to design and synthesis of small molecule libraries of biomedical importance and development of new methodologies and process developments of candidate drugs.
From left to right upper row: Dr. S.T.V.S. Kiran Kumar, Dr. Lalit Kumar, Dr. V.L. Sharma, Dr. Nand Lal, Dr. Amit Sarswat Lower row: Dhanaraju Mandalapu, Sonal Gupta, Mrs. Tara Rawat (S.T.O.), Dr. Veenu bala, Dr. Santosh Jangir
THESIS SUPERVISED
• Seven (7) students for their Ph.D. • Twenty two (22) students for their Post Graduation degrees
FORMER Ph.D. STUDENTS
• Dr. S.T.V.S. Kiran Kumar, 2006,Research Scientist at University of Virginia Charlottesville, Virginia. • Dr. Lalit Kumar, 2011, KIMIA Biosciences Pvt.Ltd., Rajasthan, India . • Dr. Amit Sarswat, 2011, Postdoctoral Fellow, University Health Network, Toronto, Ontario, Canada. • Dr. Nand Lal, 2012, Scientist E1 at HLL-Lifecare Limited, Thiruvananthapuram, Kerala, India. • Dr. Santosh Jangir, 2014. • Dr. Veenu bala, 2014, Assistant Professor at Mohan Lal Sukhdia University, Rajasthan, India. • Ms. Sonal Gupta, 2015.
FORMER PROJECT ASSISTANTS
• Ms. Mala Singh (2014-2016)
PRESENT Ph.D. STUDENTS
• Mr. Dhanaraju Mandalapu (CSIR-SRF; 2012-present)
FORMER POSTGRADUATE STUDENTS
• M. Jay Kothari (1997) • A.N. Misra (1997) • Ritu Chadda (1998) • Arun Kumar Misra (2000) • S.Nitya (2003) • Vishwanath Pratap Gupta (2004) • Divya (2006) • Charu Mahawar (2007) • Desh Deepak Pandey (2008) • Priyanka Pandey (2010) • Sumit Kumar (2010) • Sourabh Maheswari (2011) • Kartheek Nandikonda (2012) • Naveen Gupta (2012) • Pallavi Nayak (2012) • Neetika (2013) • Vikas Kumar (2013) • Neha Yadav (2013) • Subhadra Thakur (2014) • Jitendra Kumar (2015) • Suyash Tewari (2015) • Anjali Misra (2015)
MEMBERSHIP OF SOCIETES :
1. The Uttar Pradesh Association for Advancement of Science, Lucknow (India) 2. Indian Chemical Society (Calcutta) 3. Chemical Research Society of India, (Bangalore)
PROJECTS:
Reproductive Health Research: Male Reproductive Health and Contraception 1 Co – Principal Investigator: “Designed synthesis, evaluation and identification of novel, dually-effective spermicidal agents with anti-Trichomonal activity for ‘prophylactic’ contraception” (July 2014 – ongoing ), Funded by DHR, Indian council of Medical Research (ICMR), New Delhi. 2 Co-Principal Investigator: “Preclinical development of S,S’-Disulfanediylbis(pyrrolidinopropane-2,1-diyl) bis (piperidinothiocarbamate) as a vaginal contraceptive” (July 2011 – June 2013), Funded by Indian council of Medical Research (ICMR), New Delhi. 3 Principal Investigator: “Designed synthesis and biological evaluation of novel agents for management of benign prostatic hyperplasia” (November 2012 – October 2015), Funded by Indian council of Medical Research (ICMR), New Delhi.
PUBLICATIONS & PATENTS-
Total number of peer reviewed publications- 69 (Sixty Nine ) Total number of patents: (1 World patent and 4 National patents) – 5 (Five) Citations to all publications: -Sum of times cited – 486, h-index- 12
SELECTED PUBLICATIONS
• Dhanaraju Mandalapu, Deependra Kumar Singh, Sonal Gupta, Vishal M. Balaramnavar, Mohammad Shafiq, Dibyendu Banerjee, Vishnu Lal Sharma. Discovery of monocarbonyl curcumin hybrids as a novel class of human DNA ligase I inhibitors: in silico design, synthesis and biology. RSC Advances, 2016, 6, 26003. • Subhashis Pal, Kainat Khan, Shyamsundar Pal China, MonikaMittal, Konica porwal, Richa Shrivastava, Isha Taneja, Zakir Hossain, Dhanaraju Mandalapu, Jiaur R. Gayen, Muhammad Wahajuddin, Vishnu Lal Sharma, Arun K. Trivedi, Sabyasachi Sanyal, Smrati Bhadauria, Madan M. Godbole , Sushil K. Gupta, Naibedya Chattopadhyay. Theophylline, a methylxanthine drug induces osteopenia and alters calciotropic hormones and prophylactic vitamin D treatment protects against these changes in rats. Toxicology and Applied Pharmacology, 2016, 295, 12-25. • Bhavana Kushwaha, Dhanaraju Mandalapu, Veenu Bala, Lokesh Kumar, Aastha Pandey, Deepti Pandey, Santosh Kumar Yadav, Pratiksha Singh, P.K. Shukla, Jagdamba P. Maikhuri, Satya N. Sankhwar, Vishnu L. Sharma, Gopal Gupta. Ammonium salts of carbamodithioic acid as potent vaginal trichomonacides and fungicides. International Journal of Antimicrobial Agents, 2016, 47, 36-47. • Dhanaraju Mandalapu, Nand Lal, Lokesh Kumar, Bhavana Kushwaha, Sonal Gupta, Lalit Kumar, Veenu Bala, Santosh K. Yadav, Pratiksha Singh, Nidhi Singh, Jagdamba P. Maikhuri, Satya N. Sankhwar, Praveen K. Shukla, Imran Siddiqi, Gopal Gupta, Vishnu L. Sharma. Innovative Disulphide Esters of Dithiocarbamic acid as Women Controlled Contraceptive Microbicides: A Bioisosterism Approach. ChemMedChem, 2015, 10, 1739-1753. • Rachumallu Ramakrishna, Santosh kumar Puttrevu, Manisha Bhateria,Veenu Bala,Vishnu L. Sharma, Rabi Sankar Bhatta. Simultaneous determination of azilsartan and chlorthalidone in rat and human plasma by liquid chromatography-electrospray tandemmass spectrometry. Journal of Chromatography B, 2015,990, 185–197. • Hardik Chandasana, Yashpal S. Chhonkera, Veenu Bala, Yarra D. Prasad ,Telaprolu K. Chaitanya, Vishnu L. Sharma, Rabi S. Bhatta. Pharmacokinetic bioavailability, metabolism and plasma proteinbinding evaluation of NADPH-oxidase inhibitor apocynin using LC–MS/MS. Journal of Chromatography B, 2015, 985, 180–188. • Rajeev Kumar, Vikas Verma, Vikas Sharma, Ashish Jain, Vishal Singh, Amit Sarswat , Jagdamba P. Maikhuri, Vishnu L. Sharma, Gopal Gupta. A precisely substituted benzopyran targets androgen refractory prostate cancer cells through selective modulation of estrogen receptors. Toxicology and Applied Pharmacology, 2015, 283, 187-197. • Nand Lal, Amit Sarswat, Lalit Kumar, Karthik Nandikonda, Santosh Jangir, Veenu Bala, Vishnu Lal Sharma. Synthesis of Dithiocarbamates Containing Disulfide Linkage Using Cyclic Trithiocarbonate and Amines under Solvent–Catalyst Free Condition. Journal of Heterocyclic Chemistry, 2015, 52, 156-162. • Veenu Bala, Santosh Jangir, Dhanaraju Mandalapu, Sonal Gupta, Yashpal S. Chhonker, Nand Lal, Bhavana Kushwaha, Hardik Chandasana, Shagun Krishna, Kavita Rawat, Jagdamba P. Maikhuri, Rabi S. Bhatta, Mohammad I. Siddiqi,Rajkamal Tripathi, Gopal Gupta, Vishnu L. Sharma. Dithiocarbamate- Thiourea Hybrids Useful as Vaginal Microbicides Also Show Reverse Transcriptase Inhibition: Design, Synthesis, Docking and Pharmacokinetic studies. Bioorganic & Medicinal Chemistry Letters, 2015, 25, 881-886. • Gopal Gupta, Santosh Jangir and Vishnu Lal Sharma. Targeting post-ejaculation sperm for value-added contraception. Current Molecular Pharmacology, 2014, 7, 167-174. • Veenu Bala, Santosh Jangir, Vikas Kumar, Dhanaraju Mandalapu, Sonal Gupta, Lalit Kumar, Bhavana Kushwaha, Yashpal S. Chhonker, Atul Krishna, Jagdamba P. Maikhuri, Praveen K. Shukla, Rabi S. Bhatta, Gopal Gupta, Vishnu L. Sharma. Design and synthesis of substituted morpholin/piperidin-1-yl-carbamodithioates as promising vaginal microbicides with spermicidal potential. Bioorganic & Medicinal Chemistry Letters, 2014, 24, 5782-5786. • Veenu Bala, Gopal Gupta, Vishnu Lal Sharma. Chemical and Medicinal Versatility of Dithiocarbamates: An Overview. Mini Review Medicinal Chemistry, 2014, 14, 1021–1032. • Rakesh Kumar Asthana, Rasna Gupta, Nidhi Agrawal, Atul Srivastava, Upma Chaturvedi, Sanjeev Kanojiya, Ashok Kumar Khanna, Gitika Bhatia, Vishnu Lal Sharma. Evaluation of antidyslipidemic effect of mangiferin and amarogentin from swertia chirayita extract in hfd induced charles foster rat model and in vitroantioxidant activity and their docking studies. International Journal of Pharmaceutical Sciences and Research, 2014, 5(9), 3734-3740. • Santosh Jangir, Veenu Bala, Nand Lal, Lalit Kumar, Amit Sarswat, Amit Kumar, Hamidullah, Karan S. Saini, Vikas Sharma, Vikas Verma, Jagdamba P. Maikhuri, Rituraj Konwar, Gopal Gupta, Vishnu L. Sharma. Novel alkylphospholipid-DTC hybrids as promising agents against endocrine related cancers acting via modulation of Akt-pathway. European Journal of Medicinal Chemistry, 2014,85, 638-647. • Hardik Chandasana, Yashpal S. Chhonker, Veenu Bala, Yarra Durga Prasad,Vishnu L. Sharma, Rabi S. Bhatta. A rapid and sensitive LC-MS/MS analysis of diapocynin in rat plasma to investigate in vitro and in vivo pharmacokinetics.Analytical Methods 2014, 6, 7075-82. • Yashpal S. Chhonker, Hardik Chandasanaa, Veenu Bala, Lokesh Kumar,Vishnu Lal Sharma, Gopal Gupta, Rabi S. Bhatta. Quantitative determination of microbicidal spermicide ‘nonoxynol-9’ in rabbit plasma and vaginal fluid using LC–ESI–MS/MS: Application to pharmacokinetic. Journal of Chromatography B, 2014, 965, 127–132. • Mittal M, Khan K, Pal S, Porwal K, China SP, Barbhuyan TK, Bhagel KS, Rawat T, Sanyal S, Bhaduria S, Sharma VL, Chattopadhyay N. The Thiocarbamate Disulphide Drug, Disulfiram Induces Osteopenia in Rats by Inhibition of Osteoblast Function Due to Suppression of Acetaldehyde Dehydrogenase Activity.Toxicological Sciences, 2014, 239, 257-270. • Santosh Jangir, Veenu Bala, Nand Lal, Lalit Kumar, Amit Sarswat, Lokesh Kumar, Bhavana Kushwaha, Pratiksha Singh, Praveen K. Shukla, Jagdamba P. Maikhuri, Gopal Gupta, Vishnu L. Sharma. A unique dithiocarbamate chemistry during design & synthesis of novel sperm-immobilizing agents. Organic & Biomolecular Chemistry, 2014, 12 , 3090-3099. • Amit Anthwal, U. Chinna Rajesh, M.S.M. Rawat, Bhavana Kushwaha, Jagdamba P. Maikhuri, Vishnu L. Sharma, Gopal Gupta, Diwan S. Rawat. Novel metronidazole-chalcone cojugates with potential to counter drug resistance inTrichomona vaginalis. European Journal of Medicinal Chemistry, 2014, 79, 89-94. • Ashish Jain, Lokesh Kumar, Bhavana Kushwaha, Monika Sharma, Aastha Pandey, Vikas Verma, Vikas Sharma, Vishal Singh, Tara Rawat, Vishnu L. Sharma, Jagdamba P. Maikhuri, Gopal Gupta. Combining a synthetic spermicide with a natural trichomonacide for safe, prophylactic contraception. Human Reproduction, 2014, 29, 242-252. • Lalit Kumar, Nand Lal, Vikash Kumar, Amit Sarswat, Santosh Jangir, Veenu Bala, Lokesh Kumar, Bhavana Kushwaha, Atindra K. Pandey, Mohammad I. Siddiqi, Praveen K. Shukla, Jagdamba P. Maikhuri, Gopal Gupta, Vishnu L. Sharma. Azole-carbodithioate hybrids as vaginal anti-Candida contraceptive agents: design, synthesis and docking studies. European Journal of Medicinal Chemistry, 2013,70, 68-77. • Monika Sharma, Lokesh Kumar, Ashish Jain, Vikas Verma, Vikas Sharma, Bhavna Kushwaha, Nand Lal, Lalit Kumar, Tara Rawat, AK Dwivedi, JP Maikhuri, VL Sharma, Gopal Gupta. Designed chemical intervention with thiols for prophylactic contraception. PLOS-One, 2013, 8 (6), page 67365. • Lalit Kumar, Ashish Jain, Nand Lal, Amit Sarswat, Santosh Jangir, Lokesh Kumar, Priyanka Shah, Swatantra K. Jain, Jagdamba P. Maikhuri, Mohammad I. Siddiqi, Gopal Gupta, Vishnu L. Sharma. Potentiating metronidazole scaffold against resistant trichomonas: Design, synthesis, biology and 3D–QSAR analysis. ACS Medicinal Chemistry Letters, 2012, 3 (2), 83-87. • Kumar R, Verma V, Sarswat A, Maikhuri JP, Jain A, Jain RK, Sharma VL, Dalela D, Gupta G. Selective estrogen receptor modulators regulate stromal proliferation in human benign prostatic hyperplasia by multiple beneficial mechanisms-action of two new agents. Investigational New Drugs, 2012, 30, 582-593. • Ashish Jain, Nand Lal, Lokesh Kumar, Vikas Verma, Rajiv Kumar, Lalit Kumar, Vishal Singh, Raghav K. Mishra, Amit Sarswat, S. K. Jain, J. P. Maikhuri, V. L. Sharma, Gopal Gupta. Novel trichomonacidal spermicides. Antimicrobial Agents and Chemotherapy, 2011, 55 (9), 4343-4351. • Nand Lal, Lalit Kumar, Amit Sarswat, Santosh Jangir, Vishnu Lal Sharma. Synthesis of S-(2-thioxo-1,3-dithiolan-4-yl)methyl-dialkylcarbamothioate and S-thiiran-2-ylmethyl-dialkylcarbamothioate via Intermolecular O−S Rearrangement in Water. Organic Letters, 2011, 13 (9), 2330-2333. • Amit Sarswat, Rajeev Kumar, Lalit Kumar, Nand Lal, Smiriti Sharma, Yenamandra S. Prabhakar, Shailendra K. Pandey, Jawahar Lal, Vikas Verma, Ashish Jain, Jagdamba P. Maikhuri, Diwakar Dalela, Kirti, Gopal Gupta, Vishnu L. Sharma. Arylpiperazines for Management of Benign Prostatic Hyperplasia: Design, Synthesis, Quantative Structure – Activity Relationships, and Parmacokinetic Studies. Journal of Medicinal Chemistry, 2011, 54 (1), 302-311. • Lalit Kumar, Amit Sarswat, Nand Lal, Ashish Jain, Sumit Kumar, S.T.V.S. Kiran Kumar, Jagdamba P. Maikhuri, Atindra K. Pandey, Praveen K. Shukla, Gopal Gupta, Vishnu L. Sharma. Design and Synthesis of 3-(azol-1-yl)phenylprapanes as spermicide for prophylactic contraception . Bioorganic & Medicinal Chemistry Letters, 2011, 21(1), 176-181.
LIST OF PATENTS
1 Kalpana Bhandari, V.L. Sharma and S. Ray. “An improved process for the synthesis of 3,4-disubstituted-1,5-dihydro-2H-3-pyrrolin-2-one” Indian PatentAppl. 323/Del/01 dt 23.3.2001. 2 A.K.Dwivedi, V.L.Sharma, N.Kumaria, Kiran Kumar, G.Gupta, J.P.Maikhuri, J.D.Dhar, Pradeep Kumar, A.H.Ansari, P.K.Shukla, M.Kumar, Raja Roy , K.P.Madhusudanan, R.C.Gupta, Pratima Srivastava, R.Pal, and S.Singh. “Novel spermicidal and antifungal agents” Indian Patent 245815 dt 25.01.2011 ; Appl. No.1792/Del/04 dt 22.09.2004. 3 Vishanu Lal Sharma, Nand Lal, Amit Sarswat, Santosh Jangir, Veenu Bala, Lalit Kumar, Tara Rawat, Ashish Jain, Lokesh Kumar, Jagdamba Prasad Maikhuri, Gopal Gupta. “ Carbodithioates and process for preparation thereof ” NF No. 0030/NF2013/IN, Indian Patent Appl. no.0373/DEL/2013 dated 08.02.2013. 4 Vishanu Lal Sharma, Nand Lal, Amit Sarswat, Santosh Jangir, Veenu Bala, Lalit Kumar, Tara Rawat, Ashish Jain, Lokesh Kumar, Jagdamba Prasad Maikhuri, Gopal Gupta, “ Carbodithioates with spermicidal activity and process for preparation thereof ” PCT Patent no. WO 2014122670 August 14, 2014. 5 Dhanaraju Mandalapu, Rajesh K. Arigela, Tara Rawat, and Vishnu L. Sharma, “An Improved Process For Preparation Of 4-Substituted amino-2,3-polymethylenequinoline hydrochloride ” Indian Patent IN 201611003055 dated: 28.01.2016.

From left to right upper row: Dr. S.T.V.S. Kiran Kumar, Dr. Lalit Kumar, Dr. V.L. Sharma, Dr. Nand Lal, Dr. Amit Sarswat
Lower row: Dhanaraju Mandalapu, Sonal Gupta, Mrs. Tara Rawat (S.T.O.), Dr. Veenu bala, Dr. Santosh Jangir

///////////aryl piperazine, androgen sensitive prostatic disorders, 330633-91-5, CDRI-?

c1(ccc(cc1)[N+]([O-])=O)N2CCN(CC2)C(=O)CN3CCN(CC3)c4ccc(cc4)[N+]([O-])=O

DDD 107498, DDD 498

PATENT WO 2013153357,  US2015045354

6-Fluoro-2-[4-(morpholinomethyl)phenyl]-N-(2-pyrrolidin-1-ylethyl)quinoline-4-carboxamide

6-Fluoro-2-[4-(4-morpholinylmethyl)phenyl]-N-[2-(1-pyrrolidinyl)ethyl]-4-quinolinecarboxamide

4-Quinolinecarboxamide, 6-fluoro-2-[4-(4-morpholinylmethyl)phenyl]-N-[2-(1-pyrrolidinyl)ethyl]-

CAS 1469439-69-7

CAS 1469439-71-1 SUCCINATE

6-fluoro-2-[4-(morpholin-4-ylmethyl)phenyl]-N-(2-pyrrolidin-1-ylethyl)quinoline-4-carboxamide

• Originator Medicines for Malaria Venture; University of Dundee
• Class Small molecules
• Mechanism of Action Protein synthesis inhibitors

Highest Development Phases

• No development reported Malaria

Most Recent Events

• 16 Jul 2016 No recent reports of development identified for preclinical development in Malaria in United Kingdom
• 01 Apr 2015 DDD 498 licensed to Merck Serono worldwide for the treatment of Malaria

Prof Ian Gilbert:

Head of Biological Chemistry and Drug Discovery

BCDD, College of Life Sciences, University of Dundee, DD1 5EH, UK
Tel: +44 (0) 1382-386240

DDD498

Scots scientists in ‘single dose’ malaria treatment breakthrough

Scientists have discovered an antimalarial compound that could treat malaria patients in a single dose and help prevent the spread of the disease from infected people.

The compound DDD107498 also has the potential to treat patients with malaria parasites resistant to current medications, researchers say.

Scientists hope it could lead to treatments and protection against the disease, which claimed almost 600,000 lives amid 200 million reported cases in 2013.

The compound was identified through a collaboration between the University of Dundee’s drug discovery unit (DDU) and the Medicines for Malaria Venture (MMV), a separate organisation.

The compound is now undergoing further safety testing with a view to entering human clinical trials within the next year.

Details of the discovery have been published in the journal Nature.

Professor Ian Gilbert, head of chemistry at the DDU, who led the team that discovered the compound, said: “The publication describes the discovery and profiling of this exciting new compound.

“It reveals that DDD107498 has the potential to treat malaria with a single dose, prevent the spread of malaria from infected people and protect a person from developing the disease in the first place.

“There is still some way to go before the compound can be given to patients. However, we are very excited by the progress that we have made.”

The World Health Organisation reports that there were 200 million clinical cases of malaria in 2013, with 584,000 people dying from the disease. Most of these deaths were children under the age of five and pregnant women.

MMV chief executive officer Dr David Reddy said: “Malaria continues to threaten almost half of the world’s population – the half that can least afford it.

“DDD107498 is an exciting compound since it holds the promise to not only treat but also protect these vulnerable populations.

“The collaboration to identify and progress the compound, led by the drug discovery unit at the University of Dundee, drew on MMV’s network of scientists from Melbourne to San Diego.”The publication of the research is an important step and a clear testament to the power of partnership.”

MMV selected DDD107498 to enter preclinical development in October 2013 following the recommendation of its expert scientific advisory committee.

Since then, with MMV’s leadership, large quantities of the compound have been produced and it is undergoing further safety testing with a view to entering human clinical trials within the next year.

Merck Serono has recently obtained the right to develop and, if successful, commercialise the compound, with the input of MMV’s expertise in the field of malaria drug development and access and delivery in malaria-endemic countries.

Dr Michael Chew from the Wellcome Trust, which provides funding for the DDU and MMV, said: “The need for new antimalarial drugs is more urgent than ever before, with emerging strains of the parasite now showing resistance against the best available drugs.

“These strains are already present at the Myanmar-Indian border and it’s a race against time to stop resistance spreading to the most vulnerable populations in Africa.

“The discovery of this new antimalarial agent, which has shown remarkable potency against multiple stages of the malaria lifecycle, is an exciting prospect in the hunt for viable new treatments.”

PAPER

Discovery of a Quinoline-4-carboxamide Derivative with a Novel Mechanism of Action, Multistage Antimalarial Activity, and Potent in Vivo Efficacy

SCHEME 1

SCHEME 2

Preparation 4Yield: 54% Preparation 3

Yield: 27%

SCHEME 4 B

Yield: 72% Yield: 70% Preparation 6

Example 1 : 6-Fluoro-2-r4-(morpholinomethyl)phenyll-N-(2-pyrrolidin-1-ylethyl)quinoline- 4-carboxamide, Example compound 1 in Scheme 2

In a sealed microwave tube, a suspension of 2-chloro-6-fluoro-N-(2-pyrrolidin-1- ylethyl)quinoline-4-carboxamide (preparation 4) (2.00 g, 6 mmol), [4- (morpholinomethyl)phenyl]boronic acid, hydrochloride, available from UORSY, (3.20 g, 12 mmol), potassium phosphate (2.63 g, 12 mmol) and tetrakis(triphenylphosphine)palladium (0) (0.21 g, 0.19 mmol) in DMF/Water 3/1 (40 ml) was heated at 130°C under microwave irradiation for 30 min. The reaction was filtered through Celite™ and solvents were removed under reduced pressure. The resulting residue was taken up in DCM (150 ml) and washed twice with NaHC0₃ saturated aqueous solution (2 x 100 ml). The organic layer was separated, dried over MgS0₄and concentrate to dryness under reduced pressure. The reaction crude was purified by flash column chromatography using an 80 g silica gel cartridge and eluting with DCM (Solvent A) and MeOH (Solvent B) and the following gradient: 1 min hold 100% A, followed by a 30 min ramp to 10 % B, and then 15 min hold at 10% B. The fractions containing product were pooled together and concentrated to dryness under vacuum to obtain the desired product as an off-white solid (1 g). The product was dissolved in methanol (100 ml) and 3-mercaptopropyl ethyl sulfide Silica (Phosphonics, SPM-32, 60- 200 uM) was added. The suspension was stirred at room temperature over for 2 days and then at 50°C for 1 h. After cooling to room temperature, the scavenger was filtered off and washed with methanol (30 ml). The solvent was removed under reduced pressure and the product was further purified by preparative HPLC. The fractions containing product were pooled together and freeze dried to obtain the desired product as a white solid (0.6 g, 1.3 mmol, Yield 20%).

¹ H NMR (500 MHz; CDCI₃) δ 1.81-1.84 (m, 4H), 2.50-2.52 (m, 4H), 2.63 (brs, 4H), 2.82 (t, 2H, J = 5.9 Hz), 3.61 (s, 2H), 3.71 (dd, 2H, J = 5.4 Hz, J = 1 1.4 Hz), 3.74-3.76 (m, 4H), 6.84 (brs, 1 H), 7.52-7.57 (m, 3H), 7.97-8.00 (m, 2H), 8.13 (d, 2H, J = 8.2 Hz), 8.21 (dd, 1 H, J = 5.5 Hz, J = 9.2 Hz) ppm . ¹⁹ F NMR (407.5 MHz; CDCI₃) δ -11 1.47 ppm. Purity by LCMS (UV Chromatogram, 190-450nm) 99 %, rt = 5.7 min, m/z 463 (M+H)⁺ HRMS (ES+) found 463.2501 [M+H]⁺, C27H32F1 N402 requires 463.2504.

Example 2: 6-Fluoro-2-[4-(morpholinomethyl)phenyl1-N-(2-pyrrolidin-1-ylethyl)quinoline- 4-carboxamide; fumaric acid salt, compound (IB) in Scheme 2

The starting free base (example 1) (0.58 g, 1 mmol) was dissolved in dry ethanol (10 ml) and added dropwise to a stirred solution of fumaric acid (0.15 g, 1 mmol) in dry ethanol (9 ml). The mixture was stirred at room temperature for 1 h. The white precipitate was filtered, washed with ethanol (20 ml) and then dissolved in 10 ml of water and freeze dried to obtain the desired salt as a white solid (0.601 g, 1 mmol, Yield 82%).

1 H NMR (500 MHz; d6-DMSO) δ 1.83-1.86 (m, 4H), 2.41 (brs, 4H), 2.94 (brs, 4H), 3.03 (t, 2H, J = 6.2 Hz), 3.57 (s, 2H), 3.60-3.65 (m, 6H), 6.47 (s, 2H), 7.51 (d, 2H, J = 8.25), 7.74-7.78 (m, 1 H), 8.06 (dd, 1 H, J = 2.9 Hz, J = 10.4 Hz), 8.17 (dd, 1 H, J = 5.7 Hz, J = 9.3 Hz), 8.24-8.26 (m, 3H), 9.24 (t, 1 H, J = 5.5 Hz) ppm. ¹⁹ F NMR (407.5 MHz; d6- DMSO) δ -112.30 ppm.

Purity by LCMS (UV Chromatogram, 190-450nm) 99 %, rt = 5.3 min, m/z 463 (M+H)⁺

Example 1A: Alternative synthesis of 6-fluoro-2-[4-(morpholinomethyl)phenyl1-N-(2- pyrrolidin-1-ylethyl)quinoline-4-carboxamide, Example compound 1A in Scheme 4

To a stirred suspension of 6-fluoro-2-[4-(morpholinomethyl)phenyl]quinoline-4-carboxylic acid (preparation 7) (2.20 g, 6 mmol) in DCM (100 ml) at room temperature, 2-chloro- 4,6-dimethoxy-1 ,3,5-triazine (CDMT) (1.26 g, 7 mmol) and 4-methylmorpholine (NMO) (1.33 ml, 12 mmol) were added. The reaction mixture was stirred at room temperature for 1 h and then 2-pyrrolidin-1-ylethanamine (0.77 ml, 6 mmol) was added and stirred at room temperature for further 3 h. The reaction mixture was washed with NaHC0₃ saturated aqueous solution (2x 100 ml) and the organic phase was separated, dried over MgS0₄ and concentrated under reduced pressure. The resulting residue was absorbed on silica gel and purified by flash column chromatography using an 80 g silica gel cartridge and eluting with DCM (Solvent A) and MeOH (Solvent B) and the following gradient: 2 min hold 100% A followed by a 30 min ramp to 10 %B and then 15 min hold at 10%B. The desired fractions were concentrated to dryness under vacuum to obtain the crude product as a yellow solid (95% purity by LCMS). The sample was further purified by a second column chromatography using a 40 g silica gel cartridge, eluting with DCM (Solvent A) and 10% NH₃-MeOH in DCM (Solvent B) and the following gradient: 2 min hold 100% A, followed by a 10 min ramp to 23 % B and then 15 min hold at 23% B. The desired fractions were concentrated to dryness under vacuum to obtain product as a white solid (1 g). Re-crystallisation form acetonitrile (18 ml) yielded the title compound as a white solid (625 mg, 1.24 mmol, 20%).

¹ H NMR (500 MHz; CDCI₃) δ 1.81-1.84 (m, 4H), 2.50-2.52 (m, 4H), 2.63 (brs, 4H), 2.82 (t, 2H, J = 5.9 Hz), 3.61 (s, 2H), 3.71 (dd, 2H, J = 5.4 Hz, J = 1 1.4 Hz), 3.74-3.76 (m, 4H), 6.84 (brs, 1 H), 7.52-7.57 (m, 3H), 7.97-8.00 (m, 2H), 8.13 (d, 2H, J = 8.2 Hz), 8.21 (dd, 1 H, J = 5.5 Hz, J = 9.2 Hz) ppm .

¹ H NMR (500 MHz; d6-DMSO) δ 1.72-1.75 (m, 4H), 2.41 (brs, 4H), 2.56 (brs, 4H), 2.67 (t, 2H, J = 6.6 Hz), 3.49-3.52 (m, 2H), 3.56 (s, 2H), 3.60-3.61 (m, 4H), 7.52 (d, 2H, J = 8.3 Hz), 7.73-7.77 (m, 1 H), 8.07 (dd, 1 H, J = 2.9 Hz, J = 10.4 Hz), 8.18-8.21 (m, 2H), 8.26 (d, 2H , J = 8.3 Hz), 8.85 (t, 1 H, J = 6.6 Hz) ppm.

¹³C NMR (125 MHz; d⁶-DMS0₃) 5 23.2, 38.4, 53.2, 53.5, 54.5, 62.1 , 66.2, 109.0, 109.1 , 1 17.3, 120.1 , 120.3, 124.1 , 124.2, 127.1 , 129.4, 132.2, 132.3, 136.8, 139.9, 142.8, 145.2, 155.3, 159.0, 161 .0, 166.1 ppm.

¹⁹ F NM R (500 MHz; d⁶-DMSO) δ -1 12.47 ppm.

Purity by LCMS (UV Chromatogram, 190-450nm) 99 %, rt = 5.0 min, m/z 463 (M+H)⁺

Putting a stop to malaria: Phenotypic screening against malaria parasites, hit identification, and efficient lead optimization have delivered the preclinical candidate antimalarial DDD107498. This molecule is distinctive in that it has potential for use as a single-dose cure for malaria and shows a unique broad spectrum of activity against the liver, blood, and mosquito stages of the parasite life cycle.

 

Professor Ian Gilbert FRSC

Design and synthesis of potential therapeutic agents

Position:

Professor of Medicinal Chemistry and Head of the Division of Biological Chemistry and Drug Discovery

Affiliation:

Biological Chemistry and Drug Discovery

Address:

College of Life Sciences, University of Dundee, Dundee

Full Telephone:

+44 (0) 1382 386240, int ext 86240

Email:

i.h.gilbert@dundee.ac.uk

Website:

Gilbert Lab Group

Medicinal Chemistry

I am a medicinal chemist and my research interests are in the design and synthesis of potential drugs. The mainstay of my work is synthetic medicinal chemistry as part of the Drug Discovery Unit (DDU). Where possible we make extensive use of molecular modeling to guide our synthetic efforts. I have a particular interests in the following aspects of drug discovery:

Neglected diseases such as human African trypanosomiasis, leishmaniasis and malaria.
Chemical validation of drug targets, including novel targets for which there is little or no precedence for drug discovery.
Novel approaches to and paradigms for drug discovery.
Mode of action studies and target identification.

I am Head of Chemistry in the DDU. Our main focuses are on neglected diseases and novel drug targets. The neglected diseases we are tackling are malaria, tuberculosis and the kinetoplastid diseases. We use both target-based approaches and phenotypic approaches (whole parasite screening). We have had particular success in validating the enzyme N-myristoyltransferase as a drug target in human African trypanosomiasis, and in identifying and optimising phenotypic hits. In our novel targets area, we aim to validate novel areas of biology as potential drug targets.

Dr Neil Norcross

Position:

Medicinal Chemist

Affiliation:

Biological Chemistry and Drug Discovery

Address:

College of Life Sciences, University of Dundee, Dundee

Full Telephone:

(0) , int ext

Email:

n.norcross@dundee.ac.uk

///////////DDD107498, DDD 107498, PRECLINICAL, DUNDEE, MALARIA, DDD 498, Achim Porzelle, Ian Gilbert, MERCK SERENO, Beatriz Baragaña, Medicines for Malaria Venture,  University of Dundee, Neil Norcross, 1469439-69-7, 1469439-71-1 , SUCCINATE

Fc1ccc2nc(cc(c2c1)C(=O)NCCN1CCCC1)-c1ccc(cc1)CN1CCOCC1

PF-04745637

cas 1917294-46-2

MW 509.00, MF C₂₇ H₃₂ Cl F₃ N₂ O₂

Cyclopentanecarboxamide, 1-(4-chlorophenyl)-N-[2-[4-hydroxy-4-(trifluoromethyl)-1-piperidinyl]-3-phenylpropyl]-

rac-1-(4-Chlorophenyl)-N-f2-r4-hvdroxy-4-(trifluoromethyl)piperidin-1-vn-3-phenylpropyDcyclopentanecarboxamide

. Please note PF-6667294 is Compound 4 and PF-4746537 is Compound 8.

A series of potent and selective carboxamide TRPA1 antagonists were identified by a high throughput screen. Structure–activity relationship studies around this series are described, resulting in a highly potent example of the series. Pharmacokinetic and skin flux data are presented for this compound. Efficacy was observed in a topical cinnamaldehyde flare study, providing a topical proof of pharmacology for this mechanism. These data suggest TRPA1 antagonism could be a viable mechanism to treat topical conditions such as atopic dermatitis.

 

Discovery and development of TRPV1 antagonists

https://en.wikipedia.org/wiki/Discovery_and_development_of_TRPV1_antagonists

/////////////PF-04745637, PF 04745637, PF04745637, PFIZER, PRECLINICAL, TRPV1 antagonists,  atopic dermatitis, 1917294-46-2

c1(ccccc1)CC(CNC(=O)C3(c2ccc(cc2)Cl)CCCC3)N4CCC(CC4)(O)C(F)(F)F

IPI-549

CAS 1693758-51-8
MF : C30H24N8O2
Molecular Weight: 528.576

(S)-2-amino-N-(1-(8-((1-methyl-1H-pyrazol-4-yl)ethynyl)-1-oxo-2-phenyl-1,2-dihydroisoquinolin-3-yl)ethyl)pyrazolo[1,5-a]pyrimidine-3-carboxamide

2-amino-N-[(1S)-1-[8-[2-(1-methylpyrazol-4-yl)ethynyl]-1-oxo-2-phenylisoquinolin-3-yl]ethyl]pyrazolo[1,5-a]pyrimidine-3-carboxamide

• Originator Intellikine
• Developer Infinity Pharmaceuticals
• ClassAntineoplastics; Small molecules
• Mechanism of ActionPhosphatidylinositol 3 kinase delta inhibitors; Phosphatidylinositol 3 kinase gamma inhibitors

• Phase I Solid tumours

Most Recent Events

• 18 Apr 2016 Pharmacodynamics data from a preclinical study in Solid tumours presented at the 107^th Annual Meeting of the American Association for Cancer Research (AACR-2016)
• 01 Dec 2015 Phase-I clinical trials in Solid tumours (Monotherapy, Combination therapy, Late-stage disease, Second-line therapy or greater) in USA (PO)

IPI-549 is a potent and selective phosphoinositide-3-kinase (PI3Kγ) Inhibitor as an Immuno-Oncology Clinical Candidate (Kd = 0.29 nM). Bioactivity data of IPI-549: biochemcial IC50 (nM) for PI3K isoform: 3200 (α); 3500 (β); 16 (γ); and >8400 (δ) respectively. Cellar IC50 (nM) of IPI549 for PI3K isoform: 250 (α); 240 (β); 1.6 (γ); and 180 (δ) respectively. IPI-549 shows >100-fold selectivity over other lipid and protein kinases. IPI-549 demonstrates favorable pharmacokinetic properties and robust inhibition of PI3K-γ mediated neutrophil migration in vivo and is currently in Phase 1 clinical evaluation in subjects with advanced solid tumors.

PAPER

IPI-549 NMR 1H

IPI-549 13C NMR

IPI-549 ASSAY

Compound 1 is coupled to 4-ethynyl-1-methyl-1H-pyrazole using the general procedure outlined above to provide compound 26 IPI-549, in 70% yield with >98% enantiomeric purity.

IPI-549

1H NMR (400 MHz, DMSO-d6) δ 8.92 (dd, J = 6.8, 1.7 Hz, 1H), 8.55 (dd, J = 4.5, 1.7 Hz, 1H), 8.00 (d, J=6.8 Hz, 1H), 8.00 (s, 1H), 7.69 – 7.54 (m, 5H), 7.53 – 7.43 (m, 3H), 7.41 – 7.35 (m, 1H), 7.01 (dd, J = 6.7, 4.5 Hz, 1H), 6.74 (s, 1H), 6.42 (s, 2H), 4.56 (quin, J = 6.8 Hz, 1H).), 3.82 (s, 3H), 1.35 (d, J = 6.8 Hz, 3H).

13C NMR (101 MHz, DMSO-d6) δ 162.73, 161.19, 160.93, 150.06, 147.51, 146.74, 141.05, 138.09, 137.81, 135.42, 133.66, 132.56, 131.90, 129.51, 129.24, 129.20, 129.17, 128.50, 126.16, 123.41, 123.31, 107.88, 102.44, 101.15, 90.40, 87.06, 85.94, 44.88, 38.62, 20.69.

ESI-HRMS: calcd for 529.2095 C30H25N8O2 (M+H)+ , found 529.2148.

[]D 22: +447.8o (c 1.007, DMSO)

COMPD1

compound 1 in 95% yield.

1H NMR (400 MHz, CDCl3) 8.41 (dd, J = 6.8, 1.7 Hz, 1H), 8.37 (dd, J = 4.4, 1.7 Hz, 1H), 7.90 (d, J = 7.0 Hz, 1H), 7.50-7.34 (m, 5H), 7.34-7.27 (m, 2H), 6.76 (dd, J = 7.1, 4.9 Hz, 1H), 6.57 (s, 1H), 5.54 (broad s, 2H), 4.79 (quin, J = 6.9 Hz, 1H), 1.36 (d, J = 6.5 Hz, 3H);

ESI-HRMS: calcd for C24H20ClN6O2 459.1331 (M+H)+ , found 459.1386. HPLC Purity: 96% AUC.

Optimization of isoquinolinone PI3K inhibitors led to the discovery of a potent inhibitor of PI3K-γ (26 or IPI-549) with >100-fold selectivity over other lipid and protein kinases. IPI-549 demonstrates favorable pharmacokinetic properties and robust inhibition of PI3K-γ mediated neutrophil migration in vivo and is currently in Phase 1 clinical evaluation in subjects with advanced solid tumors.

Discovery of a Selective Phosphoinositide-3-Kinase (PI3K)-γ Inhibitor (IPI-549) as an Immuno-Oncology Clinical Candidate