OR

ULIXERTINIB

4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide

BVD-523; BVD-ERK; BVD-ERK/HM; BVD-ERK/ST; VRT-0752271; VRT-752271; VX-271, V

Ulixertinib malonate

4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide (referred to as ulixertinib malonate)

• Originator Vertex Pharmaceuticals
• Developer BioMed Valley Discoveries
• Class Aminopyridines; Antineoplastics; Pyrroles; Small molecules
• Mechanism of Action Mitogen activated protein kinase 3 inhibitors; Mitogen-activated protein kinase 1 inhibitor

Highest Development Phases

• Phase I/II Acute myeloid leukaemia; Cancer; Myelodysplastic syndromes
• Phase I Pancreatic cancer

Most Recent Events

• 01 Mar 2016 Phase-I clinical trials in Pancreatic cancer (Combination therapy, First-line therapy, Metastatic disease) in USA (PO) (NCT02608229)
• 23 Nov 2015 BioMed Valley Discoveries and Washington University School of Medicine plan a phase Ib trial for Pancreatic cancer (First-line therapy, Metastatic disease, Combination therapy) (PO) (NCT02608229)
• 01 Nov 2014 Phase-I/II clinical trials in Acute myeloid leukaemia (Second-line therapy or greater) and Myelodysplastic syndromes (Second-line therapy or greater) in USA (NCT02296242) (PO)

INTRODUCTION

Ulixertinib is in phase I/II clinical trials for the treatment of acute myelogenous leukemia (AML), myelodysplasia and advanced solid tumors.

Members of the family of B-cell CLL/lymphoma 2 proteins (BCL-2) are apoptosis regulators. These proteins control mitochondrial outer

membrane permeabilization (MOMP). Expression of BCL-2 protein blocks cell death in response to various cellular injuries. A number of cancers, including melanoma, breast, prostate, chronic lymphocytic leukemia, and lung cancer, may be caused by damage to the BCL-2 gene. Mutations in BCL-2 may also be a cause of resistance to cancer treatments. Unfortunately, resistance can quickly develop using conventional BCL-2 inhibitor therapies to treat cancer.

Extracellular-signal-regulated kinases (ERKs) are protein kinases that are involved in cell cycle regulation, including the regulation of meiosis, mitosis, and postmitotic functions in differentiated cells. Disruption of the ERK pathway is common in cancers. However, to date, little progress has been made developing effective ERK inhibitors for the treatment of cancer.

As the understanding of the molecular basis of cancer grows, there is an increased emphasis on developing drugs that specifically target particular nodes in pathways that lead to cancer. In view of the deficiencies noted above, there is, inter alia, a need for effective molecularly targeted cancer treatments, including combination therapies. The present invention is directed to meeting these and other needs.

Mitogen-activated protein kinase (MAPK) pathways mediate signals which control diverse cellular processes including growth, differentiation, migration, proliferation and apoptosis. One MAPK pathway, the extracellular signal-regulated kinase (ERK) signaling pathway, is often found to be up-regulated in tumors. Pathway members, therefore, represent attractive blockade targets in the development of cancer therapies (Kohno and Pouyssegur, 2006). For example, U.S. Patent No. 7,354,939 B2 discloses, inter alia, compounds effective as inhibitors of ERK protein kinase. One of these compounds, 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide, is a compound according to formula (I):

Pharmaceutical compositions are often formulated with a crystalline solid of the active pharmaceutical ingredient (API). The specific crystalline form of the API can have significant effects on properties such as stability and solubility / bioavailability. Instability and solubility characteristics can limit the ability to formulate a composition with an adequate shelf life or to effectively deliver a desired amount of a drug over a given time frame (Peterson et al., 2006).

Synergistic combination comprising an ERK1/2 inhibitor (such as ulixertinib) and a BCL-2 family inhibitor (such as navitoclax), assigned to BioMed Valley Discoveries (BVD), naming Decrescenzo and Welsch. BVD, presumably under license from Vertex, is developing ulixertinib (phase 2 trial), a small-molecule ERK 1/2 inhibitor for treating cancers including acute myelogenous leukemia and myelodysplastic syndrome. In June 2015, clinical data were presented at the 51st ASCO meeting in Chicago, IL.

BIOMED VALLEY DISCOVERIES

PATENT

WO2005113541 PDT PATENT

I-9 COMPD

SEE BELOW

PATENT

WO-2016123574

Novel crystalline forms of 4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide (referred to as ulixertinib) can be prepared which exhibit improved properties, eg surprisingly improved stability and solubility characteristics. Also claimed is their use for treating cancer.

EXAMPLE 2

Preparation of Crystaline Free Base 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide free base was prepared according to the following synthesis scheme.

Stepl

C₅H₂CIFIN

257.43 C₈H₁₀CIIN₂

ASYM-11 1606 296.54

ASYM-1 12060

ASYM-111938 ASYM-112393

ASYM-1 11935

In Step 1 , a clean and dry 200 L glass-lined reactor was evacuated to <-0.08 MPa, and then filled with nitrogen to normal pressure three times. Anhydrous ethanol (49.90 kg) was charged into the 200 L glass-lined reactor. ASYM-1 1 1606 (Asymchem) (12.70 kg) and isopropylamine (29.00 kg) were added into the mixture in turn. The mixture was heated to 65-75°C for refluxing. The mixture reacted at 65-75°C. After 20 h, the reaction was sampled and analyzed by HPLC every 4-6 h until the content of ASYM-1 1 1606 was <1 %. The mixture was cooled to 40-45°C and was concentrated at <45°C under reduced pressure (<-0.08 MPa) until 13-26 Lremained. The organic phase was washed with a sodium chloride solution and was stirred for 20-30 min and settled for 20-30 min before separation. The organic phase was concentrated at <30°C under reduced pressure (<-0.06 MPa) until 13-20 L remained. Petroleum ether (8.55 kg) was added into the concentrated mixture. The mixture was transferred into a 20 L rotary evaporator and continued concentrating at <30°C under reduced pressure (<-0.06 MPa) until 13-20 L remained. Then petroleum ether (8.55 kg) was added into the concentrated mixture. The mixture was cooled to 0-5°C and stirred for crystallization. After 1 h, the mixture was sampled and analyzed by wt% every 1 -2 h until the wt% of the mother liquor was <1 1 % or the change of the wt% between consecutive samples was <1 %. The mixture was filtered with a 10 L filter flask. The filter cake was sampled and analyzed for purity by HPLC. 10.50 kg of product was recovered as a brownish yellow solid at 99.39% purity.

In Step 2, a clean and dry 300 L glass-lined reactor was evacuated to <-0.08 MPa, and then filled with nitrogen to normal pressure three times. Glycol dimethyl ether (73.10 kg) was charged into the 300 L glass-lined reactor at 20-30°C. ASYM-1 12060 (Asymchem) (10.46 kg) and ASYM-1 1 1938 (Asymchem) (12.34 kg, 1 1 .64 kg after corrected) were added into the mixture in turn under the protection of nitrogen. Maintaining the temperature at 20-30°C, purified water (10.50 kg) and anhydrous sodium carbonate (5.67 kg) were added into the mixture. Palladium acetate (0.239 kg) and tricyclohexylphosphonium tetrafluoroborate (0.522 kg) were added into the mixture under the protection of nitrogen. After addition, the mixture was evacuated to <-0.06 MPa, and then filled with nitrogen to normal pressure. This was repeated for ten times until residual oxygen was <300 ppm. The mixture was heated to 75-85°C for refluxing. The mixture reacted at 75-85°C. After 4 h, the mixture was sampled and analyzed by HPLC every 2-3 h for content of ASYM-

1 12060. The content of AS YM-1 12060 was 6.18%, so additional ASYM-1 1 1938 (0.72 kg) was added and continued reaction until the content of ASYM-1 12060 was <3%. The mixture was cooled to 25-35°C and filtered with a 30 L stainless steel vacuum filter. The filter cake was soaked and washed twice with THF (14.10kg). The filtrate and washing liquor were combined and concentrated at <50°C under reduced pressure (<-0.08 MPa) until 10-15 L remained. The mixture was cooled to 15-25°C. Methanol (1 1 .05 kg) was added into the concentrated mixture. Then the mixture was stirred for crystallization. After 2 h, the mixture was sampled and analyzed by HPLC every 2-4 h until the wt% of the mother liquor was <2%. The mixture was filtered with a 30 L stainless steel vacuum filter. The filter cake was soaked and washed twice with methanol (8.30 kg). The filter cake was transferred into a 50 L plastic drum. Then ethyl acetate (7.10 kg) and petroleum ether (46.30 kg) were added into the drum. The mixture was stirred for 1.5-2 h and then filtered with a nutsche filter. The filter cake was soaked and washed with petroleum ether (20.50 kg). The filter cake was dried in the nutsche filter under nitrogen at 30-40°C. After 8 h, the solid was sampled and Karl Fischer (KF) analysis was performed in intervals of 4-8 h to monitor the drying process. Drying was completed when the KF result was <1 .0% water. During drying, the solid was turned over and mixed every 4-6 h. 12.15 kg of product was recovered as a brownish yellow solid at 98.32% purity.

In Step 3, a clean and dry 300 L glass-lined reactor was evacuated to <-0.08 MPa, and then filled with nitrogen to normal pressure three times. THF (62.58 kg) was charged into the 300 L glass-lined reactor at 15-30°C. Then the stirrer was started. ASYM-1 12393 (12.00 kg, 1 1 .70 kg after corrected) was added into the mixture. The mixture was stirred until the solid dissolved completely. Maintaining the temperature at 15-30°C, a lithium hydroxide solution which was

prepared with lithium hydroxide monohydrate (5.50 kg) in purified water (70.28 kg) was added into the mixture. Then diethylamine (3.86 kg) was added. The mixture was heated to 60-70°C for refluxing. The mixture reacted at 60-70°C. After 30 h, the reaction was sampled and analyzed by HPLC every 4-6 h until the content of intermediate at relative retention time (RRT)=1 .39-1 .44 was <1 % and the content of ASYM-1 12393 was <1 %. HPLC conditions for this analysis are set forth in Table 1 .

Table 1 : HPLC Parameters

The mixture was cooled to 25-35°C and MTBE (25.97 kg) was added into the mixture. The mixture was stirred for 20-30 min and filtered via an in-line fluid filter. The filtrate was transferred into a 300 L glass-lined reactor and settled for 20-30 min before separation. The pH of the obtained aqueous phase was adjusted with a 6 N hydrochloric acid solution which was prepared from concentrated hydrochloric acid (14.86 kg) in purified water (10.88 kg) at the rate of 5-8 kg/h at 15-25°C until the pH was 1 -2. The pH of the mixture was adjusted again with a saturated sodium carbonate solution which was prepared from sodium carbonate (5.03 kg) in purified water (23.56 kg) at the rate of 3-5 kg/h at 15-25°C until the pH was 6.4-6.7. Then the pH of the mixture was adjusted with a hydrochloric acid solution which was prepared from concentrated hydrochloric acid (1 .09 kg) in purified water (0.80 kg) until the pH was 6.2-6.4. The mixture was filtered with a nutsche filter. The filter cake was transferred into a 300 L glass-lined reactor and purified water (1 17.00 kg) was added. The mixture was stirred and sampled and analyzed by HPLC until the p-toluenesulfonic acid residue of the filter cake was <0.5%. Then the mixture was filtered. The filter cake was dried in the tray drier under nitrogen at 55-65°C until KF<10%. The solid and MTBE (8.81 kg) were charged into a 50 L stainless steel drum. The mixture was stirred for 1 -2 h. The mixture was filtered with a 30 L stainless steel vacuum filter. The filter cake was dried in the nutsche filter at 50-60°C. After 8 h, the solid was sampled and analyzed by KF every 4-8 h until KF<5%. During drying, the solid was turned over and mixed every 4-6 h. 6.3 kg of product was recovered as an off-white solid at 98.07% purity.

In Step 4, a dry and clean 50 L flask was purged with nitrogen for 20 min. DMF (30.20 kg) was charged into the 50 L flask reactor. Then the stirrer was started. Maintaining the temperature at 15-25°C, ASYM-1 12394 (3.22 kg, 2.76 kg after corrected) was added into the mixture. The mixture was stirred until the solid dissolved completely. The mixture was cooled to -10 to -20°C and 1 -hydroxybenzotriazole hydrate (2.10 kg) was added into the mixture at -10 to -20°C. Then EDCI (2.41 kg) was added into the mixture in five portions at an interval of about 5-10 min. The mixture was cooled to -20 to -30°C and ASYM-1 1 1888 (Asymchem) (1 .96 kg) was added into the mixture at -20 to -30°C. Then DIEA (1 .77 kg) was added into the mixture at the rate of 3-4 kg/h. The mixture was heated to 15-25°C at the rate of 5-10°C/h. The mixture was reacted at 15-25°C. After 6-8 h, the mixture was sampled and analyzed by HPLC every 2-4 h until the content of ASYM-1 12394 was <2%. The mixture was cooled to 0-10°C and the reaction mixture was quenched with a solution which was prepared from ethyl acetate (28.80 kg) in purified water (12.80 kg) at 0-10°C. The mixture was extracted three times with ethyl acetate (28.80 kg). For each extraction the mixture was stirred for 20-30 min and settled for 20-30 min before separation. The organic phases were combined and washed twice with purified water (12.80 kg). The mixture was stirred for 20-30 min and settled for 20-30 min before separation for each time. Then the obtained organic phase was filtered through an in-line fluid filter. The filtrate was transferred into a 300 L glass-lined reactor. The mixture was washed twice with a 5% acetic acid solution, which was prepared from acetic acid (2.24 kg) in purified water (42.50 kg). The solution was added at the rate of 10-20 kg/h. The organic phase was washed twice with a sodium carbonate solution, which was prepared from sodium carbonate (9.41 kg) in purified water (48.00 kg). The organic phase was washed twice with a sodium chloride solution, which was prepared from sodium chloride (16.00 kg) in purified water (44.80 kg). The organic phase was transferred into a 300 L glass-lined reactor. Anhydrous sodium sulfate (9.70 kg) was added into the mixture and the mixture was stirred for 2-4 h at 15-30°C. The mixture was filtered with a nutsche filter, which was pre-loaded with about 1 cm thick silica gel (7.50 kg). The filter cake was soaked and washed with ethyl acetate (14.40 kg) before filtration. The filtrates were combined and the combined filtrate was added into a 72 L flask through an in-line fluid filter. The mixture was concentrated at T≤40°C under reduced pressure (P<-0.08 MPa) until 3-4 L remained. Then MTBE (4.78 kg) was added into the mixture. The mixture was cooled to 0-10°C for crystallization with stirring. After 1 h, the mixture was sampled and analyzed by wt% every 1-2 h until the wt% of the mother liquor was <5% or the change of wt% between consecutive samples was <1%. The mixture was filtered with a vacuum filter flask and the filter cake was dried in the tray drier under nitrogen at 30-40°C until KF<0.5%. 3.55 kg of product was recovered as an off-white solid at 100% purity.

EXAMPLE 3A

Preparation of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide Form C

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide Form C was prepared from 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide free base as follows. ASYM-1 1 1935 (10.4 kg) was added to a stirred mixture of anhydrous ethanol (73.9 kg), methanol (4.1 kg) and isopropanol (4.1 kg). The mixture was heated to 70-75°C and stirred until all the solids dissolved. Anhydrous HCI (37 wt%, 1 .1 eq) in a mixture of ethanol/methanol/isopropanol (90:5:5) was added and the mixture maintained at 70-75°C for 2 hours after the addition was completed. The mixture was then cooled to 15-25°C at a rate of 5-15°C per hour and stirred at this temperature until the desired polymorphic purity was reached. The end point of the crystallization/polymorph conversion was

determined by the absence of an XRPD peak at about 10.5° 2Θ in three successive samples.

The mixture was then filtered and washed successively with a pre-prepared solution of anhydrous ethanol (14.8 kg), methanol (0.8 kg) and isopropanol (0.8 kg), followed by MTBE (2 x 21 kg). Avoidance of delay in the washing of the filter cake is preferable because the polymorph may be unstable in the wet filter cake in the presence of reagent alcohol and improved stability was observed after the MTBE wash has been performed. The wet filter cake was then dried in a heated filter funnel or a tray drier at 40-50°C until dry. Typical yields were about 85-90%.

EXAMPLE 3B

Alternative Preparation of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide Form C

ASYM-1 15985

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide Form C was also prepared from 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide free base as follows. A dry and clean 72 L flask was purged with nitrogen for 20 min. Anhydrous ethanol (21 .35 kg) methanol (1 .17 kg) and isopropanol (1 .19 kg) were charged into the 72 L flask at 15-25°C and the mixture was stirred for 20-30 min. ASYM-1 1 1935 (3.01 kg) was added into the mixture and heated to 70-75°C at the rate of 15-25°C/h and stirred until the solid dissolved completely.

An alcohol / HCI solution was prepared as follows. Anhydrous ethanol (1.500 kg) methanol (0.088 kg) and isopropanol (0.087 kg) were charged into a 5 L flask at 15-25°C and the mixture was stirred for 20-30 min. The mixture was bubbled with hydrogen chloride through a dip tube under stirring at 10-25°C. After 2 h, the mixture was sampled and analyzed every 2-4 h until the wt% of hydrogen chloride was > 35%.

The alcohol / HCI solution (0.519 kg) prepared above was added dropwise into the mixture at the rate of 0.5-1.0 kg/h at 70-75°C. Seed crystal (0.009 kg) was added into the mixture and the alcohol / HCI solution (0.173 kg) prepared above was added into the mixture at the rate of 0.5-1 .0 kg/h at 70-75°C. After addition, the mixture was stirred for 1 -2 h at 70-75°C. The mixture was cooled to 15-25°C at the rate of 5-15°C/h and stirred for 4-6 h. The mixture was heated to 70-75°C at the rate of 15-25°C/h and stirred for 8-10 h at 70-75°C. The mixture was cooled to 15-25°C at the rate of 5-15°C/h and stirred for 4-6 h. The mixture was filtered with a vacuum filter flask. The filter cake was soaked and rinsed with a solution which was prepared from anhydrous ethanol (4.25 kg) and methanol (0.24 kg) and isopropanol (0.24 kg) before filtration. The filter cake was dried in a drying room under nitrogen at 40-50°C until the ethanol residue was <0.5% and methanol residue was <0.3% and isopropanol residue was <0.3%. 2.89 kg of product was recovered as a white solid at 99.97% purity.

PATENT

WO-2016123581

Novel crystalline malonate salt forms of 4-(5-chloro-2-isopropylaminopyridin-4-yl)-1H-pyrrole-2-carboxylic acid[1-(3-chlorophenyl)-2-hydroxyethyl]amide (referred to as ulixertinib malonate) and composition comprising them. Also claimed is their use for treating cancer.

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2016123581&redirectedID=true

EXAMPLE 6

Aqueous Disolution of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1-(3-chlorophenyl)-2-hydroxyethyl]amide Malonate Form A

Samples of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide Form C and 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2 -carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide malonate Form A were each shaken at ambient temperature in fasting state simulated gastric fluid (FaSSGF) pH 1.6 for 30 minutes. Concentration of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide was measured at 5, 15 and 30 minutes.

After 30 minutes, the samples were removed from FaSSGF, placed in fasting state simulated intestinal fluid (FaSSIF) pH 6.5, with shaking, for an additional 5 hours. Concentration of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide was measured at 10, 30, 60 90, 120, 180, 270, and 300 minutes. Results are summarized in Table 13 and shown in FIG. 10A (FaSSGF) and FIG. 10B (FaSSIF).

Table 13: Solubility of 4-(5-Chloro-2-isopropylaminopyridin-4-yl)-1 H-pyrrole-2-carboxylic acid [1 -(3-chlorophenyl)-2-hydroxyethyl]amide Form C and Malonate Form A.

PATENT

WO2016123574

PATENT

WO2015095834

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2015095834&redirectedID=true

PATENT

WO2005113541

Example 1 Compound 1-9 was prepared as follows:

1-9

2,2,2-TrichIoro-l-(4-iodo-lH-pyrrol-2-yl)ethanone: To a stirred solution of 50 g (235 mmol, 1.0 equiv.) of 2,2,2-trichloro-l-(lH-pyrrol-2-yl)-ethanone in dry dichloromethane (400 mL) under nitrogen, a solution of iodine monochloride (39 g, 240 mmol, 1.02 equivalents) in of dichloromethane (200 mL) was added dropwise. The resulting mixture was stirred at room temperature for 2 hours. The solution was washed with 10% potassium carbonate, water, 1.0 M sodium thiosulfate, saturated sodium chloride, dried over sodium sulfate, filtered, and concentrated under reduced pressure. The solid was recrystallized from hexanes/methyl acetate to afford the title compound (68.5g, 86%) as a colorless solid (86%). MS FIA: 335.8, 337.8 ES-.

4-Iodo-lH-pyrrole-2-carboxyIic acid methyl ester: To a stirred solution of 2,2,2- trichloro-l-(4-iodo-lH-pyrrol-2-yl)ethanone (68g, 201 mmol, 1.0 equivalent) in dry methanol (400 mL) under nitrogen, was added a solution of sodium methoxide in methanol (4.37 M, 54 mL, 235 mmol, 1.2 equivalents) over 10 minutes. The resulting mixture was stirred at room temperature for 1 hour. The volatiles were removed under reduced pressure and the crude was then partitioned between water and tert- butylmethyl ether. The organic phase was separated, washed two times with water, saturated sodium chloride, dried over sodium sulfate, filtered and concentrated under vacuum to afford the title compound (48g, 96%) as a colorless solid, that was used directly without further purification.

4-Iodo-l-(toluene-4-sulfonyl)-lH-pyrrole-2-carboxylic acid methyl ester: 4-Iodo- lH-pyrrole-2-carboxylic acid methyl ester (24.6 g, 98 mmol, 1.0 equivalent) was dissolved in dichloromethane (150 mL) and triethylamine (30 mL, 215.6 mmol, 2.2 equivalents). 4-(Dimethylamino)pyridine (1.2 g, 9.8 mmol, 0.1 equivalent) and p- toluenesulfonylchloride (20.6 g, 107.8 mmol, 1.1 equivalents) were added and the reaction mixture was stirred for 16 hours at room temperature. The reaction was quenched with 1 M ΗC1 and the organic layer was washed with aqueous sodium bicarbonate and brine. After drying over magnesium sulfate, the solvent was removed under reduced pressure and the residue was crystallized from tert-butylmethyl ether, yielding the title compound as a pale yellow solid (30 g, 75%). R_t(min) 8.259 minutes.

4-(4,4,5,5-Tetramethyl-[l,3,2]dioxaborolan-2-yI)-l-(toluene-4-sulfonyl)-lH- pyrrole-2-carboxylic acid methyl ester: To a degassed solution of 4-iodo-l- (toluene-4-sulfonyl)-lH-pyrrole-2-carboxylic acid methyl ester (20 g, 49.4 mmol, 1.0 equivalent) and bis(pinacolato)diborane (15 g, 65 mmol, 1.3 equivalents) in DMF (200 mL) under nitrogen, was added dichloro[l,l ‘- bis(diphenylphosphino)ferrocene]palladium (II) dichloromethane adduct (3.6 g, 4.9 mmol, 0.1 equivalent). The reaction mixture was then stirred at 80 °C for 18 hours. After removing the DMF under reduced pressure, the resulting thick oil residue was suspended in diethyl ether (500 mL) and a solid precipitated immediately. This solid was removed by filtration and the filtrate was washed with IM HCl, water, brine and dried over MgS0 . Concentration afforded the title compound as a white solid and used without further purification (10 g, 50%). LC/MS: R_t(min) 4.6; 406.4 ES+. MS FIA: 406.2 ES+. ‘pfNMR δ 1.2 (s, 12H), 2.35 (s, 3H), 3.8 (s, 3H), 7.2 (m, 3H), 7.8 (d, 2H), 8.0 (s, IH).

N,N’-2-(5-Chloro-4-iodo-pyridyI)-isopropyIarnine:

Method A. (Microwave)

In a 10 mL microwave tube, 5-chloro-2-fluoro-4-iodopyridine (1.0 g, 3.9 mmol, 1.0 equivalent) was dissolved in DMSO (4.0 mL) and then ispropylamine (0.99 mL, 11.7 mmol, 3.0 equivalents) was added. The tube was sealed and placed under microwave irradiation for 600 sec at 150 °C. This reaction was repeated six times. The reaction mixtures were combined, then diluted in ethyl acetate and washed with water. After drying over sodium sulfate, the solvent was evaporated to afford the title compound as a thick brown oil (5.6 g, 80% ) which was used directly without further purification. R_t(min) 4.614; MS FIA: 296.9 ES+. ‘pfNMRsssssss δ 1.25 (d, 6H), 3.65 (m, IH), 7.15 (s, IH), 7.75 (s, IH).

Method B: (Thennal)

5-Chloro-2-fluoro-4-iodopyridine (400 mg, 1.55 mmol, 1.0 equivalent) was dissolved in ethanol (5.0 mL) and then isopropylamine (0.66 mL, 7.8 mmol, 5.0 equivalents) was added. The resulting solution was stirred at 80 °C for 48 hours. The reaction mixture was then diluted in ethyl acetate and washed with water. After drying over sodium sulfate, the solvent was evaporated and a thick brown oil was obtained, which was then purified by flash chromatography on silica gel eluting with mixtures of hexanes/ethyl acetate (from 99:1 to 80:20) to afford the title compound as a pale yellow solid (96 mg, 21%).

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-l-(toluene-4-suIfonyl)-lH-pyrrole-2- carboxylic acid methyl ester: To a solution of N,N’-2-(5-chloro-4-iodo-pyridyl)- isopropylamine (0.53 g, 1.8 mmol, 1.0 equivalent) and 4-(4,4,5,5-tetramethyl- [l,3,2]dioxaborolan-2-yl)-l-(toluene-4-sulfonyl)-lH-pyrrole-2-carboxylic acid methyl ester (0.78 g, 1.8 mmol, 1.0 equivalent) in DME (4.0 mL) was added a solution of aqueous 2 M sodium carbonate (1.0 mL) followed by Pd(PPh₃)₄ (0.21 mg, 0.18 mmol, 0.1 equivalent). The microwave tube was sealed and the reaction mixture was irradiated by microwave for 1800 sec. at 170 °C. The cmde of six reactions were combined and diluted in ethyl acetate and washed with water. After drying the organic layer with sodium sulfate, the solvent was removed and the resulting thick oil was adsorbed on silica gel. The crude was then purified by flash chromatography on silica, eluting with hexanes/ethyl acetate mixtures (from 99:1 to 70:30) to afford the title compound as a yellow solid (3.1 g, 61% over two steps). R_t(min) 6.556. MS FIA: 448.1 ES+. ‘HNMR δ 1.45 (d, 6H), 2.5 (s, 3H), 3.81 (s, 3H), 6.8 (s, IH), 7.35 (s, IH),

7.4 (d, 2H), 8.0 (m ,3H), 8.3 (s, IH).

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-l-(2,4,6-trimethylbenzenesulfonyl)- lH-pyrrole-2-carboxylic acid methyl ester: To a solution of N,N’-2-(5-chloro-4- iodo-pyridyl)-isopropylamine (96 mg, 0.32 mmol, 1.0 equivalent) and 4-(4,4,5,5- tetramethyl-[ 1 ,3,2]dioxaborolan-2-yl)- 1 -(2,4,6-trimethylbenzenesulfonyl)- lH-pyrrole- 2-carboxylic acid methyl ester (152 mg, 0.35 mmol, 1.1 equivalents) in DME (2 mL), was added a solution of aqueous 2 M sodium carbonate (0.2 mL) followed by Pd(PPh ) (37 mg, 0.032 mmol, 0.1 equivalent). The reaction mixture was stirred at 80 °C for 16 hours. The crude was diluted in ethyl acetate and washed with water. After drying the organic layer with sodium sulfate, the solvent was removed and the resulting thick oil was adsorbed on silica gel. The cmde was then purified by flash chromatography on silica, eluting with hexanes/ethyl acetate mixtures (from 99:1 to 80:20) to afford the title compound as a yellow solid (65 mg, 43%). R_t(min) 7.290. MS FIA:476.1 ES+.

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxyIic acid:

Method A. (Microwave)

A solution of 4-(5-chloro-2-isopropylaminopyridin-4-yl)-l-(toluene-4-sulfonyl)-lH- pyrrole-2-carboxylic acid methyl ester (3.1 g, 6.9 mmol, 1.0 equivalent) in TΗF (2.0 mL) was added to a solution of lithium hydroxide monohydrated (710 mg, 17.3 mmol,

2.5 equivalents) in water (3.0 mL). The microwave tube was sealed and the reaction mixture was irradiated by microwave for 1200 sec. at 150 °C. The cmde solution was acidified with aqueous 6Ν ΗC1. The solvent was evaporated off to afford the title compound which was used directly without further purification. R_t(min): 3.574. FIA MS: 279.9 ES+; 278.2 ES-.

Method B: (Thermal)

A solution of 4-(5-chloro-2-isopropylaminoρyridin-4-yl)-l-(2,4,6- trimethylbenzenesulfonyl)-lH-pyrrole-2-carboxylic acid methyl ester (0.69 g, 1.4 mmol, 1.0 equivalent) in TΗF (3.0 mL) was added to a solution of lithium hydroxide monohydrated (1.19 g, 29 mmol, 20.0 equivalents) in water (3.0 mL). The mixture was then refluxed for 8 hours. The cmde solution was acidified with aqueous 6N ΗC1 until cloudy, the organic solvent was partially removed and the product precipitated. The title compound was isolated by filtration and washed with water and diethyl ether, yielding a white solid (0.38 g, 96%).

4-(5-Chloro-2-isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxyIic acid [l-(3- ch!orophenyl)-2-hydroxyethyl] amide: To a suspension of 4-(5-chloro-2- isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid (1.93 g, 6.9 mmol, 1.0 equivalent) in DMF (5.0 mL) was added EDCI (1.45 g, 7.6 mmol, 1.1 equivalents), ΗOBt (0.94 g, 6.9 mmol, 1.0 equivalent) and (5)-3-chlorophenylglycynol (1.58 g, 7.6 mmol, 1.1 equivalents). Dusopropylethylamme (2.7 mL) was then added and the resulting mixture was stirred a room temperature overnight. The mixture was then poured into water and extracted with ethyl acetate. After drying over sodium sulfate, the solvent was removed and the crude was adsorbed on silica gel. Purification was effected by flash chromatography on silica, eluting with mixtures of hexanes/acetone (from 80:20 to 60:40) to afford the title compound as white solid (1.9 g, 64%). R_t(min) 4.981s. FIA MS: 433.1 ES+; 431.2 ES-. ¹ΗNMR (CD₃OD) δ 1.31 (d, 6H), 3.85 (m, 3H), 5.15 (t, IH), 7.01 (s, IH), 7.25 (m, 3H), 7.4 (s, IH), 7.45 (s, IH), 7.7 (s, IH), 7.95 (s, IH).

Example 2 Compound 1-9 was also prepared according to following alternate method:

2,5-DichIoro-4-nitropyridine N-oxide: To a suspension of 2-chloro-5-chloropyridine (10 g, 0.067 mol) in acetic anhydride (25 mL) was added hydrogen peroxide 30% (25 mL) in small portions. This mixture was stirred at room temperature for 24 hours and then heated at 60 °C for 30 hours. After removing the excess of acetic acid under reduced pressure, the residue was added in small portions to concentrated sulfuric acid (15 mL). The resulting solution was added to a mixture of concentrated sulfuric acid (15 mL) and fuming nitric acid (25 mL) and then heated at 100 °C for 90 minutes. The reaction mixture was poured on ice, neutralized with solid ammonium carbonate and finally with aqueous ammonia until a basic pH was obtained and. A precipitate formed. The precipitate was collected by filtration to afford the title compound as a pale yellow solid (3.1 g), R_t(min) 3.75. MS FIA shows no peak. ‘pfΝMR (DMSO-de) δ 8.78 (s, IH), 9.15 (s, IH).

4-Bromo-2-chloro-5-N-isopropylpyridin-2-amine N-oxide: To 2,5-dichloro-4- nitropyridine Ν-oxide (400 mg, 1.9 mmol) was added acetyl bromide (2 mL) very slowly. The reaction mixture was then heated at 80 °C for 10 minutes. The solvent was removed under a stream of nitrogen and the cmde product was dried under high vacuum. The cmde material (165 mg, 0.62 mmol) was dissolved in ethanol (2 mL), z^‘so-propylamine (0.53 mL) added and the resulting mixture was heated at 80 °C for 2 hours. The cmde solution was then purified by reversed phase HPLC (acetonitrile/water/TFA 1%) to afford the title compound as a pale yellow solid (60 mg, 36.6%). R_t(min) 5.275. MS FIA264.8, 266.9 ES+.

4-(5-chloro-2-isopropylaminopyridin-4-yl)-lH-pyrrole-2-carboxylic acid [l-(3- chlorophenyl)-2-hydroxyethyl] amide (1-9): 4-Bromo-2-chloro-5-N- isopropylpyridin-2-amine N-oxide (25 mg, 0.094 mmol, 1.0 equivalent) and 4- (4,4,5, 5-tetramethyl-[l,3,2]dioxaborolan-2-yl)-l-(2,4,6-trimethylbenzensulfonyl)-lH- pyrrole-2-carboxylic acid methyl ester (39 mg, 0.094 mmol, 1.0 equivalent) were dissolved in benzene (5 mL) then aqueous 2M Νa₂C0₃ (1 mL) and Pd(PPh₃)₄ (115.6 mg, 0.1 mmol, 0.2 equivalent) were added and the resulting suspension was heated at reflux at 80 °C for 16 hours. The reaction mixture was diluted in ethyl acetate, washed with water and dried over anhydrous sodium sulfate to afford 4-(5-chloro-2- isopropylamino-pyridin-4-yl)- 1 -(2,4,6-trimethyl-benzenesulfonyl)- lH-pyrrole-2- carboxylic acid methyl ester N-oxide (R (min) 6.859. MS FIA: 492.0 ES+) which was then treated with a 2 M solution of PC1₃ in dichloromethane (1 mL) at room temperature. After 10 minutes, the solvent was removed under a stream of nitrogen and the cmde oil was dissolved in methanol (1 mL) and aqueous 1 M ΝaOΗ (1 mL). The resulting mixture was heated at reflux for 16 hours then the cmde solution was acidified using aqueous 1 M ΗC1 and the solvent was removed. The resulting 4-(5- chloro-2-isopropylamino-pyridin-4-yl)-lΗ-pyrrole-2-carboxylic acid (R (min) 3.527. MS FIA: 279.4 ES+; 278.2 Es-) was suspended in DMF (3 mL) together with EDCI (36 mg, 0.19 mmol, 2 equivalents), HOBt (26 mg, 0.19 mmol, 2 equivalents), (S)-3- chlorophenylglycinol HCl salt (59 mg, 0.28 mmol, 3 equivalents) and DIEA (0.12 mL, 0.75 mmol, 8 equivalents). The resulting mixture was stirred at room temperature for 16 hours. The reaction mixture was diluted in ethyl acetate, washed with water and dried over sodium sulfate. After removing the solvent under reduced pressure, the cmde product was purified by reversed phase HPLC (acetonitrile/water/TFA 1%) to afford the title compound as a white solid (4.8 mg, 8.1%).

PATENT

US20150512092015-02-19COMPOUNDS AND COMPOSITIONS AS INHIBITORS OF MEK

US73549392008-04-08Pyrrole inhibitors of ERK protein kinase, synthesis thereof and intermediates thereto

Research scientist Tony Huang works in a laboratory at Vertex Pharmaceuticals Inc. in San Diego

REFERENCES

1 . Kohno M, Pouyssegur J (2006) Targeting the ERK signaling pathway in cancer therapy. Ann Med 38: 200-21 1 .

2. Kuby, J., Immunology, 3rd Ed., W.H. Freeman & Co., New York.

3. Lee DC, Webb ML(2003) Pharmaceutical Analysis. John Wiley & Sons, Inc., New York: 255-257.

4. Peterson ML, Hickey MB, Zaworotko MJ and Almarsson O (2006) Expanding the Scope of Crystal Form Evaluation in Pharmaceutical Science. J Pharm Pharmaceut Sci 9(3):317-326.

5. Pierce Catalog and Handbook, 1994-1995; Pierce Chemical Co., Rockford, III.

6. Remington, The Science and Practice of Pharmacy (21 st Edition, Lippincott Williams and Wilkins, Philadelphia, PA.

7. The United States Pharmacopeia-National Formulary, The United States Pharmacopeial Convention, Rockville, MD.

Gabriel Martinez-Botella

///////////ULIXERTINIB, BVD-523; BVD-ERK,  BVD-ERK/HM,  BVD-ERK/ST,  VRT-0752271,  VRT-752271,  VX-271, уликсертиниб ,أوليكسيرتينيب  ,优立替尼 , PHASE 2,  Vertex Pharmaceuticals, BioMed Valley Discoveries, UNII:16ZDH50O1U,  869886-67-9 , Gabriel Martinez-Botella

CC(C)NC1=NC=C(C(=C1)C2=CNC(=C2)C(=O)NC(CO)C3=CC(=CC=C3)Cl)Cl

Piramal Enterprises Limited announced that its wholly owned subsidiary in the U.S. has entered into an agreement to acquire 100 percent stake in Ash Stevens Inc., a U.S.-based contract development and manufacturing organization (CDMO), in an all cash deal for a consideration of USD $42.95 million plus an earn-out consideration capped at $10 million. This potential transaction is expected to be completed by the end of August.

Located in Riverview, Michigan, Ash Stevens has over 50 years of experience in contract manufacturing, and serves several biotech, mid-size pharma, and large pharmaceutical clients worldwide.

With over 60,000 sq. ft. of facilities, eight chemical drug development and production laboratories, and six full-scale production areas, Ash Stevens has built a stellar reputation, led by science, driven by operational excellence, and one that emphasizes quality as a culture. As one of the leaders in HPAPI manufacture, Ash Stevens has an impeccable safety record of working with high potency anti-cancer agents and other highly-potent therapeutics. The state-of-the-art manufacturing facility in Michigan features all necessary engineering and containment controls for the safe handling and cGMP manufacture of small and large-scale HPAPIs, with Occupational Exposure Limits (OELs) ≤ 0.1µg/m3. The facility has approvals from U.S., EU, Australia, Japan, Korea, and Mexico regulatory agencies.

“The acquisition of Ash Stevens fits well with our strategy to build an asset platform that offers value to our partners and collaborators. Currently, around 25 percent of the molecules in clinical development are potent. Our clients are looking for reliable partners that can assist them in advancing these programs forward,” said Vivek Sharma, CEO of Piramal Pharma Solutions. He further adds, “North America is a key market that we can now service with our three local facilities – the Coldstream Labs in Kentucky for fill finish needs, the Torcan facility in Toronto for complex high value APIs and now, Ash Stevens in Michigan for HPAPIs. Having facilities with a differentiated platform and geographical proximity to clients are keys towards building strategic partnerships. We expect this acquisition to also be synergistic with our Antibody Drug Conjugates (ADCs) and injectable business. We can now fulfill client requirements for a single source of supply for both high potent APIs and drug products.”

“With its rich history of scientific excellence, a track record of 12 product launches, Ash Stevens is well poised to become the partner of choice for clients looking to advance programs from early development through launch. In addition to the business benefits that the combined entity will bring to our clients, I am also pleased that the firms share common core values: both were founded by successful entrepreneurs, value integrity, and are committed to a customer-first approach,” said Dr. Mark Cassidy, President of the API Business at Piramal Pharma Solutions. “I am pleased to welcome the Ash Stevens team into the Piramal group. We expect them to be an integral part of our future growth plans.”

Added Dr. Stephen Munk, CEO of Ash Stevens, “We look forward to working with the Piramal leadership and management team, to develop API solutions that benefit customers and improve the lives of patients. The commitment that Piramal has shown towards growing its healthcare businesses, coupled with the complementary capabilities that our two firms have, makes this an exciting time for Ash Stevens and our employees. We have already identified areas where we can create significant value together, and will be moving forward rapidly to achieve those objectives.”

The transaction is not subject to any regulatory approvals. No related party of PEL has any interest in Ash Stevens.

Wells Fargo Securities, LLC served as exclusive financial advisor to Ash Stevens, with legal counsel provided by Morrison & Foerster LLP.

For further information on the financials, please visit our website: www.piramal.com.

Dr. Stephen A. Munk, President and CEO of Ash Stevens Inc.

Large scale reactor train with 2000, 3000, and 4000 L glass-lined reactors equipped with split butterfly valves.

Ash Stevens’ down draft kilo suite with low temperature capability.

Ajay Piramal

////////////CDMO,  Ash Stevens, Piramal Enterprises, Stephen A. Munk

Anders Eriksson, Matti Lepistö, Michael Lundkvist, af Rosenschöld Magnus Munck,Pavol Zlatoidsky,

Astrazeneca Ab INNOVATOR

• OriginatorAstraZeneca
• Class
• Mechanism of ActionMatrix metalloproteinase inhibitors
• Highest Development Phases

• DiscontinuedChronic obstructive pulmonary disease

Most Recent Events

• 29 Jul 2010Discontinued – Phase-II for Chronic obstructive pulmonary disease in Europe (PO)
• 29 Jul 2010Discontinued – Phase-I for Chronic obstructive pulmonary disease in Japan (PO)
• 29 Jul 2010Discontinued – Phase-I for Chronic obstructive pulmonary disease in Japan (PO)

AZD1236 is a selective MMP-9 and MMP-12 inhibitor (IC50 4.5 and 6.1nM) from Astrazeneca that, since it failed biomarker endpoints for COPD is included in the AZ Open Innovation 2014 set for repurposing. Pending any published link the structure identification is tenatative but seems likely to be the structure crystalised in WO2007106022.

Matrix metallopeptidase 9 and 12 (MMP9|MMP12) inhibitor http://www.ncbi.nlm.nih.gov/gene/4318; http://www.ncbi.nlm.nih.gov/gene/4321 Preclinical Pharmacology AZD1236 is a potent and reversible inhibitor of human MMP9 and MMP12 (IC50’s = 4.5 and 6.1nM, respectively), with 10 – 15-fold selectivity to MMP2 and MMP13 and >350-fold selectivity to other members of the enzyme family. Its activity is approximately 20- to 50-fold lower at the rat, mouse, and guinea pig orthologues. In acute models of lung injury, AZD1236 inhibited the hemorrhage and inflammation induced by instillation of human MMP12 into rat lungs by ~80% at 0.81 mg/kg, and also abolished macrophage infiltration into BAL fluid induced by tobacco smoke inhalation in the mouse. Safety and Tolerability In healthy human volunteers, AZD1236 was well tolerated in single doses from 2 to 1500 mg and in multiple doses of 15, 75 and 500 mg for periods of up to 13 days. AZD1236 was also well tolerated in COPD patients with moderate to severe disease when given at 75 mg BID for 6 weeks. Pre-clinical toxicology studies of up to 12 month duration have been performed. Toxicologically important findings mainly relate to chronic treatment and included: diffuse eye lens opacities after 6 months administration to rats and fibrodysplasia in the subcutis after 12 months to dogs. Clinical Pharmacology Target coverage data to date have been mixed. In healthy subjects, single dose of 30 or 75 mg inhibited ex vivo zymosanstimmulated whole blood MMP activity (the 75 mg dose yielding plasma compound levels at Cmax steady state of ~120 x IC50). In contrast, 75 mg BID for 6 wks in COPD patients compared to placebo did not identify any significant change in whole blood MMP activity.

PATENT

WO 2002074750 

WO 02/074767 further discloses a specific metalloproteinase inhibitor compound identified therein as (5S)-5-[4-(5-chloro-pyridin-2-yloxy)-piperidine-l-sulfonylmethyl]-5- methyl-imidazolidine-2,4-dione (page 65, lines 15 to 27; and page 120, lines 23 to 29). This compound is designated herein as compound (I).

(I)

WO 02/074767 further discloses processes for the preparation of compound (I). Thus, in one embodiment, compound (I) is prepared by a route analogous to that shown in the following Scheme (WO 02/074767; pages 87, 113 and 120) but substituting the appropriate amine in step (d):

Scheme 1

Reagents and conditions for Scheme 1: a) KCN, (NHLj)₂CO₃, EtOHTH₂O, +90⁰C, 3h;. b) Chiral separation, CHIRALPAK AD, methanol as eluent;. c) Cl₂ (g), AcOH/H₂O, <+15 ⁰C, 25min; d) Diisopropylethylamine, THF. -20 ⁰C, 30 min.

The obtained compound (I) is then purified either by precipitation and washing with ethanol/water or by preparative HPLC. In a second embodiment, the racemate of compound (I), (5RS)-5-[4-(5-chloro-pyridin-2- yloxy)-piperidine-l-sulfonylmethyl]-5-methyl-imidazolidine-2,4-dione, was prepared by reacting l-[4-(5-chloro-pyridin-2-yloxy)-piperidine-l-sulfonyl]-propan-2-one with an excess of potassium cyanide and ammonium carbonate in ethanol, and isolating the product by precipitation. Compound (I), the (5S)-enantiomer, was then obtained by chiral HPLC (WO 02/074767; pages 55 and 65).

No crystalline forms of compound (I) are disclosed in WO 02/074767.

Compound (I) is a potent metalloproteinase inhibitor, particularly a potent inhibitor of

MMP 12, and as such is useful in therapy. However, when made according to the processes described in WO 02/074767, compound (I) exhibits unpredictable solid state properties with respect to thermodynamic stability. To prepare pharmaceutical formulations containing compound (I) for administration to humans in accordance with the requirements of U.S. and other international health registration authorities, there is a need to produce compound (I) in a stable form, such as a stable crystalline form, having constant physical properties.

PATENT

WO  2007106022

WO 02/074767 further discloses a specific metalloproteinase inhibitor compound identified therein as (5S)-5-[4-(5-chloro-pyridin-2-yloxy)-piperidine-l-sulfonylmethyl]-5- methyl-imidazolidine-2,4-dione (page 65, lines 15 to 27; and page 120, lines 23 to 29). This compound is designated herein as compound (I).

(I)

WO 02/074767 further discloses processes for the preparation of compound (I). Thus, in one embodiment, compound (I) is prepared by a route analogous to that shown in the following Scheme (WO 02/074767; pages 87, 113 and 120) but substituting the appropriate amine in step (d):

Scheme 1

Reagents and conditions for Scheme 1: a) KCN, (NHLj)₂CO₃, EtOHTH₂O, +90⁰C, 3h;. b) Chiral separation, CHIRALPAK AD, methanol as eluent;. c) Cl₂ (g), AcOH/H₂O, <+15 ⁰C, 25min; d) Diisopropylethylamine, THF. -20 ⁰C, 30 min.

The obtained compound (I) is then purified either by precipitation and washing with ethanol/water or by preparative HPLC. In a second embodiment, the racemate of compound (I), (5RS)-5-[4-(5-chloro-pyridin-2- yloxy)-piperidine-l-sulfonylmethyl]-5-methyl-imidazolidine-2,4-dione, was prepared by reacting l-[4-(5-chloro-pyridin-2-yloxy)-piperidine-l-sulfonyl]-propan-2-one with an excess of potassium cyanide and ammonium carbonate in ethanol, and isolating the product by precipitation. Compound (I), the (5S)-enantiomer, was then obtained by chiral HPLC (WO 02/074767; pages 55 and 65).

No crystalline forms of compound (I) are disclosed in WO 02/074767.

Compound (I) is a potent metalloproteinase inhibitor, particularly a potent inhibitor of

MMP 12, and as such is useful in therapy. However, when made according to the processes described in WO 02/074767, compound (I) exhibits unpredictable solid state properties with respect to thermodynamic stability. To prepare pharmaceutical formulations containing compound (I) for administration to humans in accordance with the requirements of U.S. and other international health registration authorities, there is a need to produce compound (I) in a stable form, such as a stable crystalline form, having constant physical properties.

A preferred process for the synthesis of compound (I) is shown in Scheme 2.

Scheme 2

KCN, (NH₄)₂CO₃

(H) 2-propanol

Chromatography KOBu’

Cl₂

AcOH AcOH, H₂O

Compound (I)

Recrystallisation EtOH, H₂O

Compound (I) Form G

Example 5

(S)-5-[4-(5-ChIoro-pyridin-2-yloxy)-piperidine-l-suIfonylmethyl]-5-methyl- imidazoIidine-2,4-dione Process 1

I₅ a) 5-Chloro-2-(piperidin-4-yloxy)-pyridine

5-Chloro-2-(piperidin-4-yloxy)-pyridine acetate (40 g, 0.146 mol) was slurried in iso- PrOAc (664 mL) at 30⁰C. To this slurry was added Na₂CO₃ (1.5 mol per litre; 196 mL, 2 mol eq.). The slurry was then rapidly stirred at 30 ⁰C for 15 minutes. The biphasic mixture was allowed to settle, and the bottom aqueous phase was separated and discarded.

20 The above base washing procedure was repeated twice more. The organic phase was then washed once with water (200 mL). The resulting iso-VxOAc solution was reduced in volume to approximately 300 mL by distillation under reduced pressure. The solution was then diluted with zsø-PrOAc (400 mL) and again distilled down to approximately 300 mL. This procedure was repeated once more. A sample was removed for analysis of 5-chloro-

25 2-(piperidm-4-yloxy)-pyridine content and water content. The weight or the volume of the solution was measured in order to calculate the concentration of 5-chloro-2-(piperidin-4- yloxy)-pyridme in the Z-PrOAc solution.

fr) rSV5-r4-(5-Chloro-pyridin-2-yloxyVpiperidine-l-sulfonylmethvn-5-methyl- 30 imidazolidine-2 ,4-dione Diisopropylethylamine (24.3 mL, 0.139 mol, 1 mol eq.) was added to the iso-PrOAc solution prepared in part (a) [ca. 300 mL; equivalent to 31.2 g, 0.146 mol, 1.05 mol eq. of 5-chloro-2-(piperidin-4-yloxy)-pyridine] in one portion at RT. The solution was then cooled to -15 °C.

((S)-4-Methyl-2,5-dioxo-imidazolidin-4-yl)-methanesulfonyl chloride (31.65 g, 0.139 mol, 1 mol eq.) was dissolved in dry THF (285 mL) at RT with stirring. The resulting solution was then added to the iso-PrOAc solution of 5-chloro-2-(piperidin-4-yloxy)- pyridine dropwise at -15 ⁰C over about 1.5 h. A precipitate was seen on addition of the ((S)-4-methyl-2,5-dioxo-imidazolidin-4-yl)-methanesulfonyl chloride. At the end of the addition, dry THF (32 mL) was added to the reaction mixture to wash the line and the mixture was stirred for 1 h at — 15 ⁰C. It was then warmed to 20 °C over 1 h and stirred at 20 °C for 1 h further. The reaction was quenched with 10 wt% NaHSO₄ (157 mL) with rapid stirring. After about 15 minutes, the biphasic mixture was allowed to settle, and the bottom aqueous phase was separated and discarded. This acid wash procedure was repeated once more. The organic phase was then washed with water (157 mL) using rapid stirring and allowing complete phase separation before partitioning. The reaction solution was then warmed to 40 °C and washed again with water (157 mL). THF (95 mL) was added to the organic layer that was then warmed to 40 ⁰C and filtered at 40⁰C to remove any particulate matter. The solvent volume was then reduced to about 157 mL by reduced pressure distillation with the jacket temperature at 55 °C. zso-PrOAc (317 mL) was then added and the volume was again reduced to about 157 mL. Two more put-and-takes of zsø-PrOAc (317 mL) were carried out. Solids began to precipitate out during the distillations and a suspension resulted. The volume was reduced to about 157 mL each time and after the final distillation a small sample of solvent was then removed from the reaction mixture for residual THF analysis. The ¹H NMR showed no THF peaks. The contents of the reaction were then cooled to 0 °C and the product was collected by filtration. The reaction vessel was washed with zsø-PrOAc (63 mL) and this rinse was used to wash the product on the filter. The product was dried overnight in a vacuum oven at 40 °C. The required (S)-5-[4- (5-chloro-pyridin-2-yloxy)-piperidine-l-sulfonyhnethyl]-5-methyl-imidazolidine-2,4-dione was isolated as a white solid in 71% yield (41.8 g).

¹H NMR (300MHz, d6-DMSO) δ 10.74 (IH, s), 8.20 (IH, d), 8.01 (IH, s), 7.81 (IH, dd), 6.87 (IH, d), 5.09 (IH, m), 3.52-3.35 (4H, m), 3.13 (2H, m), 2.02 (2H, m), 1.72 (2H, m), 1.33 (3H, s).

Example 6

(S)-5-[4-(5-Chloro-pyridin-2-yloxy)-piperidine-l-sulfonylmethyl]-5-methyl- imidazolidine-2,4-dione Process 2 a) 5-Chloro-2-(piperidm-4-yloxy)-pyridine

5-Chloro-2-(piperidm-4-yloxy)-pyridine acetate (70 g, 257 mmol) was slurried in toluene

(560 mL) at RT. IM NaOH (420 mL) was added and the slurry was then rapidly stirred at RT for 15 min. The biphasic mixture was allowed to settle, and the bottom aqueous phase was separated and discarded. The organic phase was then washed with water (2 x 420 mL). A sample was removed from the organic phase and assayed for 5-chloro-2-(piperidin-

4-yloxy)-pyridine.

The resulting toluene solution was then reduced in volume by distillation at reduced pressure, down to approximately 168 mL (2.4 vol eq. with respect to 5-chloro-2-(piperidin-

4-yloxy)-pyridine acetate charge). The solution was then diluted with toluene (420 mL) and again distilled down to approx 168 mL (2.4 vol eq.). A sample was removed for analysis of water content.

b^*) (S)-5-r4-r5-Chloro-pyridm-2-yloxy)-piperidine-l-sulfonylmethvH-5-methyl- imidazolidine-2 ,4-dione

Diisopropylethylamine (38.4 mL, 220 mmol) was added to the toluene solution of 5-chloro-2-(piperidin-4-yloxy)-pyridine obtained in step (a) (containing 236 mmol) in one portion followed by dry THF (151 mL) as a line wash. ((S)-4-Methyl-2,5-dioxo- imidazolidin-4-yl)-methanesulfonyl chloride (48.7 g, 215 mmol) was dissolved in dry THF (352 mL) at RT with stirring. The resulting solution of the sulfonyl chloride was then added dropwise to the toluene/THF solution of 5-chloro-2-(piperidin-4-yloxy)-pyridine and diisopropylethylamine at RT over 1 to 2 h. A precipitate was seen on addition of the sulfonyl chloride. At the end of the addition, dry THF (50 mL) was added to the reaction 5 mixture as a line wash. After the addition was complete, the reaction was stirred for about 30 min at RT.

The reaction was quenched with 10 wt% NaHSO₄ (251 mL) with rapid stirring for approx 15 min. The biphasic mixture was allowed to settle, when the bottom aqueous phase was io separated and discarded. This acid wash procedure was repeated once more. The solvent volume was then reduced to about 220 mL by reduced pressure distillation. Toluene (300 mL) was then added and the volume was reduced to about 245 mL Solids begin to precipitate during the distillations and a suspension resulted. After the final distillation, a small sample of solvent was then removed from the reaction mixture for residual THF i₅ analysis.

The contents of the reaction mixture were then cooled to 0 °C, stirred for about 30 minutes at this temperature and the product was collected by filtration. The reaction vessel was washed with toluene (100 mL) and this rinse was used to wash the product on the filter. 20 The product was dried in a vacuum oven at 40 ⁰C to constant weight. (S)-5-[4-(5-Chloro- pyridin-2-yloxy)-piperidine-l-sulfonylmethyl]-5-methyl-imidazolidine-2,4-dione was isolated as a white solid in typically 85 to 88% yield over the two steps.

Aerial view of Mölndal

Patent

WO 2003106689

Paul Hudson, President, AstraZeneca U.S. and Executive Vice President, North America, joined by AstraZeneca volunteers to celebrate the AstraZeneca Hope Lodge’s fifth birthday.

CLIPS

Astra boss Pascal Soriot

Massachusetts Economic Development Secretary Jay Ash (left) congratulates Kumar Srinivasan, Head of AstraZeneca R&D Boston (right), at a ceremony to launch AstraZeneca’s Gatehouse Park BioHub.

https://ncats.nih.gov/files/AZD1236.pdf

///////AZD1236,  AZD-1236, AZD 1236,

O=S(=O)(C[C@@]1(C)NC(=O)NC1=O)N3CCC(Oc2ccc(Cl)cn2)CC3

Osanetant (SR-142,801)

160492-56-8 CAS

(R)-(+)-N-[[3-[1-benzoyl-3-(3,4-dichlorophenyl)piperidin-3-yl]prop-1-yl]-4-phenylpiperidin-4-yl]-N-methylacetamide

Osanetant (SR-142,801) was a neurokinin 3 receptor antagonist developed by Sanofi-Synthélabo, which was being researched for the treatment of schizophrenia, but was discontinued.^[1][2] It was the first non-peptide NK₃ antagonist developed in the mid-1990s,^[3][4] Other potential applications for osanetant is in the treatment of drug addiction, as it has been found to block the effects ofcocaine in animal models.^[5][6]

Developed by Sanofi-Aventis (formerly Sanofi-Synthelabo), osanetant (SR-142801) is an NK3 receptor antagonist which was under development for the treatment of schizophrenia and other Central Nervous System (CNS) disorders. In a review of its R&D portfolio, the company announced in August 2005 that it would cease any further development ofosanetant. This follows an earlier decision to discontinue development of eplivanserin for schizophrenia

References

1.  “osanetant Sanofi-Aventis discontinued, France.”. Highbeam.
2. Kamali, F (July 2001). “Osanetant Sanofi-Synthélabo”. Current opinion in investigational drugs (London, England : 2000). 2 (7): 950–6.PMID 11757797.
3.  Emonds-Alt, X; Bichon, D; Ducoux, JP; Heaulme, M; Miloux, B; Poncelet, M; Proietto, V; Van Broeck, D; et al. (1995). “SR 142801, the first potent non-peptide antagonist of the tachykinin NK3 receptor”. Life Sciences. 56 (1): PL27–32. doi:10.1016/0024-3205(94)00413-M.PMID 7830490.
4.  Quartara L, Altamura M (August 2006). “Tachykinin receptors antagonists: from research to clinic”. Current Drug Targets. 7 (8): 975–92.doi:10.2174/138945006778019381. PMID 16918326. Retrieved 2011-04-14.
5.  Desouzasilva, M; Mellojr, E; Muller, C; Jocham, G; Maior, R; Huston, J; Tomaz, C; Barros, M (May 2006). “The tachykinin NK3 receptor antagonist SR142801 blocks the behavioral effects of cocaine in marmoset monkeys”. European Journal of Pharmacology. 536 (3): 269–78.doi:10.1016/j.ejphar.2006.03.010. PMID 16603151.
6.  Jocham, Gerhard; Lezoch, Katharina; Müller, Christian P.; Kart-Teke, Emriye; Huston, Joseph P.; De Souza Silva, M. AngéLica (September 2006). “Neurokinin receptor antagonism attenuates cocaine’s behavioural activating effects yet potentiates its dopamine-enhancing action in the nucleus accumbens core”. European Journal of Neuroscience. 24 (6): 1721–32. doi:10.1111/j.1460-9568.2006.05041.x.PMID 17004936.

Osanetant

Systematic (IUPAC) name
N-(1-{3-[(3R)-1-benzoyl-3-(3,4-dichlorophenyl)piperidin-3-yl]propyl}-4-phenylpiperidin-4-yl]-N-methylacetamide
Identifiers
CAS Number	160492-56-8
ATC code	none
PubChem	CID 219077
IUPHAR/BPS	2110
ChemSpider	189901
UNII	K7G81N94DT
ChEMBL	CHEMBL346178
Chemical data
Formula	C35H41Cl2N3O2
Molar mass	606.625 g/mol

///////Osanetant , SR-142,801, 

CC(=O)N(C)C1(CCN(CC1)CCC[C@@]2(CCCN(C2)C(=O)C3=CC=CC=C3)C4=CC(=C(C=C4)Cl)Cl)C5=CC=CC=C5

LINK

http://drugapprovalsint.com/

http://drugapprovalsint.com/

http://drugapprovalsint.com/

http://drugapprovalsint.com/

/////////////

ACT-334441

Cenerimod

UNII-Y333RS1786; Y333RS1786

S1P receptor 1 agonist

CAS 1262414-04-9
Chemical Formula: C25H31N3O5
Exact Mass: 453.22637

Actelion Pharmaceuticals Ltd.

Martin Bolli, Cyrille Lescop, Boris Mathys,Keith Morrison, Claus Mueller, Oliver Nayler,Beat Steiner,

(S)-3-(4-(5-(2-cyclopentyl-6-methoxypyridin-4-yl)-1,2,4-oxadiazol-3-yl)-2-ethyl-6-methylphenoxy)propane-1,2-diol

(2S)-3-[4-[5-(2-cyclopentyl-6-methoxypyridin-4-yl)-1,2,4-oxadiazol-3-yl]-2-ethyl-6-methylphenoxy]propane-1,2-diol

(S)-3-{4-[5-(2-Cyclopentyl-6-methoxy-pyridin-4-yl)-[1,2,4]oxadiazol-3-yl]-2-ethyl-6-methyl-phenoxy}-propane-1,2-diol

Cenerimod is a potent and orally active immunomodulator, exhibited EC50 value of 2.7 nM. Cenerimod is an agonist for the G protein-coupled receptor S1 P1/EDG1 and has a powerful and long-lasting immunomodulating effect which is achieved by reducing the number of circulating and infiltrating T- and B-lymphocytes, without affecting their maturation, memory, or expansion. Cenerimod may be useful for prevention or treatment of diseases associated with an activated immune system

CENERIMOD

ACT-334441; lysosphingolipid receptor agonist – Actelion; S1P₁ receptor modulator – Actelion; Second selective S1P1 receptor agonist – Actelion; Sphingosine 1 phosphate receptor modulators – Actelion; Sphingosine 1-phosphate receptor 1 agonists – Actelion

• Mechanism of Action Lysosphingolipid receptor agonists

• Highest Development Phases

• Phase I/II Systemic lupus erythematosus

Most Recent Events

• 09 Jun 2016 Actelion terminates a phase I drug interaction trial for Systemic lupus erythematosus (In volunteers) in France (NCT02479204)
• 22 Dec 2015 Phase-I/II clinical trials in Systemic lupus erythematosus in Ukraine, Belarus (PO) (NCT02472795)
• 24 Sep 2015 Phase-I/II clinical trials in Systemic lupus erythematosus in USA (PO) (NCT02472795)

The human immune system is designed to defend the body against foreign micro-organisms and substances that cause infection or disease. Complex regulatory mechanisms ensure that the immune response is targeted against the intruding substance or organism and not against the host. In some cases, these control mechanisms are unregulated and autoimmune responses can develop. A consequence of the uncontrolled inflammatory response is severe organ, cell, tissue or joint damage. With current treatment, the whole immune system is usually suppressed and the body’s ability to react to infections is also severely compromised. Typical drugs in this class include azathioprine, chlorambucil, cyclophosphamide, cyclosporin, or methotrexate. Corticosteroids which reduce inflammation and suppress the immune response, may cause side effects when used in long term treatment. Nonsteroidal anti-inflammatory drugs (NSAIDs) can reduce pain and inflammation, however, they exhibit considerable side effects. Alternative treatments include agents that activate or block cytokine signaling.

Orally active compounds with immunomodulating properties, without compromising immune responses and with reduced side effects would significantly improve current treatments of uncontrolled inflammatory diseases.

In the field of organ transplantation the host immune response must be suppressed to prevent organ rejection. Organ transplant recipients can experience some rejection even when they are taking immunosuppressive drugs. Rejection occurs most frequently in the first few weeks after transplantation, but rejection episodes can also happen months or even years after transplantation. Combinations of up to three or four medications are commonly used to give maximum protection against rejection while minimizing side effects. Current standard drugs used to treat the rejection of transplanted organs interfere with discrete intracellular pathways in the activation of T-type or B-type white blood cells. Examples of such drugs are cyclosporin, daclizumab, basiliximab, everolimus, or FK506, which interfere with cytokine release or signaling; azathioprine or leflunomide, which inhibit nucleotide synthesis; or 15-deoxyspergualin, an inhibitor of leukocyte differentiation.

The beneficial effects of broad immunosuppressive therapies relate to their effects; however, the generalized immunosuppression which these drugs produce diminishes the immune system’s defense against infection and malignancies. Furthermore, standard immunosuppressive drugs are often used at high dosages and can cause or accelerate organ damage.

SYNTHESIS

PATENT

https://www.google.com/patents/WO2011007324A1?cl=zh

The human immune system is designed to defend the body against foreign microorganisms and substances that cause infection or disease. Complex regulatory mechanisms ensure that the immune response is targeted against the intruding substance or organism and not against the host. In some cases, these control mechanisms are unregulated and autoimmune responses can develop. A consequence of the uncontrolled inflammatory response is severe organ, cell, tissue or joint damage. With current treatment, the whole immune system is usually suppressed and the body’s ability to react to infections is also severely compromised. Typical drugs in this class include azathioprine, chlorambucil, cyclophosphamide, cyclosporin, or methotrexate. Corticosteroids which reduce inflammation and suppress the immune response, may cause side effects when used in long term treatment. Nonsteroidal anti-inflammatory drugs (NSAIDs) can reduce pain and inflammation, however, they exhibit considerable side effects. Alternative treatments include agents that activate or block cytokine signaling.

Orally active compounds with immunomodulating properties, without compromising immune responses and with reduced side effects would significantly improve current treatments of uncontrolled inflammatory diseases.

In the field of organ transplantation the host immune response must be suppressed to prevent organ rejection. Organ transplant recipients can experience some rejection even when they are taking immunosuppressive drugs. Rejection occurs most frequently in the first few weeks after transplantation, but rejection episodes can also happen months or even years after transplantation. Combinations of up to three or four medications are commonly used to give maximum protection against rejection while minimizing side effects. Current standard drugs used to treat the rejection of transplanted organs interfere with discrete intracellular pathways in the activation of T-type or B-type white blood cells. Examples of such drugs are cyclosporin, daclizumab, basiliximab, everolimus, or FK506, which interfere with cytokine release or signaling; azathioprine or leflunomide, which inhibit nucleotide synthesis; or 15-deoxyspergualin, an inhibitor of leukocyte differentiation.

The beneficial effects of broad immunosuppressive therapies relate to their effects; however, the generalized immunosuppression which these drugs produce diminishes the immune system’s defense against infection and malignancies. Furthermore, standard immunosuppressive drugs are often used at high dosages and can cause or accelerate organ damage.

Description of the invention

The present invention provides novel compounds of Formula (I) that are agonists for the G protein-coupled receptor S1 P1/EDG1 and have a powerful and long-lasting immunomodulating effect which is achieved by reducing the number of circulating and infiltrating T- and B-lymphocytes, without affecting their maturation, memory, or expansion. The reduction of circulating T- / B-lymphocytes as a result of S1 P1/EDG1 agonism, possibly in combination with the observed improvement of endothelial cell layer function associated with S1 P1/EDG1 activation, makes such compounds useful to treat uncontrolled inflammatory diseases and to improve vascular functionality. Prior art document WO 2008/029371 discloses compounds that act as S1 P1/EDG1 receptor agonists and show an immunomodulating effect as described above. Unexpectedly, it has been found that the compounds of the present invention have a reduced potential to constrict airway tissue/vessels when compared to compounds of the prior art document WO 2008/029371. The compounds of the present invention therefore demonstrate superiority with respect to their safety profile, e.g. a lower risk of bronchoconstriction.

Examples of WO 2008/029371 , which are considered closest prior art analogues are shown in Figure 1.

Figure 1 : Structure of Examples of prior art document WO 2008/029371 , which are considered closest analogues to the compounds of the present invention.

The data on the constriction of rat trachea rings compiled in Table 1 illustrate the superiority of the compounds of the present invention as compared to compounds of prior art document WO 2008/029371.

For instance, the compounds of Example 1 and 6 of the present invention show a significantly reduced potential to constrict rat trachea rings when compared to the compounds of prior art Examples 222 and 226 of WO 2008/029371 , respectively. Furthermore, the compounds of Example 1 and 6 of the present invention also show a reduced potential to constrict rat trachea rings when compared to the compounds of prior art Examples 196 and 204 of WO 2008/029371 , respectively. These data demonstrate that compounds wherein R¹ represents 3-pentyl and R² represents methoxy are superior compared to the closest prior art compounds of WO 2008/029371 , i.e. the compounds wherein R¹ represents an isobutyl and R² represents methoxy or wherein R¹represents methyl and R² represents 3-pentyl. Moreover, also the compound of Example 16 of the present invention, wherein R¹ is 3-methyl-but-1-yl and R² is methoxy, exhibits a markedly reduced potential to constrict rat trachea rings when compared to its closest analogue prior art Example 226 of WO 2008/029371 wherein R¹ is isobutyl and R² is methoxy.

The unexpected superiority of the compounds of the present invention is also evident from the observation that the compounds of Example 2 and 7 of the present invention show a markedly reduced potential to constrict rat trachea rings when compared to the compounds of prior art Examples 229 and 233 of WO 2008/029371 , respectively. This proves that compounds wherein R¹represents cyclopentyl and R² represents methoxy are superior compared to the closest prior art compounds of WO 2008/029371 , i.e. the compounds wherein R¹ represents methyl and R² represents cyclopentyl.

Also, the compound of Example 3 of the present invention exhibits the same low potential to constrict rat trachea rings as its S-enantiomer, i.e. the compound of Example 2 of the present invention, indicating that the configuration at this position has no significant effect on trachea constriction. Furthermore, also Example 21 of the present invention exhibits the same low potential to constrict rat trachea rings as present Example 2, which differs from Example 21 only by the linker A (forming a 5-pyridin-4-yl-[1 ,2,4]oxadiazole instead of a 3- pyridin-4-yl-[1 ,2,4]oxadiazole). This indicates that also the nature of the oxadiazole is not critical regarding trachea constriction.

Table 1 : Rat trachea constriction in % of the constriction induced by 50 mM KCI. n.d. = not determined. For experimental details and further data see Example 33.

result obtained at a compound concentration of 300 nM.

The compounds of the present invention can be utilized alone or in combination with standard drugs inhibiting T-cell activation, to provide a new immunomodulating therapy with a reduced propensity for infections when compared to standard immunosuppressive therapy. Furthermore, the compounds of the present invention can be used in combination with reduced dosages of traditional immunosuppressant therapies, to provide on the one hand effective immunomodulating activity, while on the other hand reducing end organ damage associated with higher doses of standard immunosuppressive drugs. The observation of improved endothelial cell layer function associated with S1 P1/EDG1 activation provides additional benefits of compounds to improve vascular function.

The nucleotide sequence and the amino acid sequence for the human S1 P1/EDG1 receptor are known in the art and are published in e.g.: HIa, T., and Maciag, T., J. Biol

Chem. 265 (1990), 9308-9313; WO 91/15583 published 17 October 1991 ; WO 99/46277 published 16 September 1999. The potency and efficacy of the compounds of Formula (I) are assessed using a GTPγS assay to determine EC₅O values and by measuring the circulating lymphocytes in the rat after oral administration, respectively (see in experimental part). i) In a first embodiment, the invention relates to pyridine compounds of the Formula (I),

Formula (I)

PATENT

WO 2013175397

https://www.google.com/patents/WO2013175397A1?cl=en

Pyridine-4-yl derivatives of formula (PD),

Formula (PD) A represents

(the asterisks indicate the bond that is linked to the pyridine group of Formula (PD));

R^a represents 3-pentyl, 3-methyl-but-1-yl, cyclopentyl, or cyclohexyl;

R^b represents methoxy;

R^c represents 2,3-dihydroxypropoxy, -OCH₂-CH(OH)-CH₂-NHCO-CH₂OH,

-OCH₂-CH(OH)-CH₂N(CH₃)-CO-CH₂OH, -NHS0₂CH₃, or -NHS0₂CH₂CH₃; and

R^d represents ethyl or chloro.)

disclosed in WO201 1007324, have immunomodulating activity through their S1 P1/EDG1 receptor agonistic activity. Therefore, those pyridine-4-yl derivatives are useful for prevention and / or treatment of diseases or disorders associated with an activated immune system, including rejection of transplanted organs such as kidney, liver, heart, lung, pancreas, cornea, and skin; graft-versus-host diseases brought about by stem cell transplantation; autoimmune syndromes including rheumatoid arthritis, multiple sclerosis, inflammatory bowel diseases such as Crohn’s disease and ulcerative colitis, psoriasis, psoriatic arthritis, thyroiditis such as Hashimoto’s thyroiditis, uveo-retinitis; atopic diseases such as rhinitis, conjunctivitis, dermatitis; asthma; type I diabetes; post-infectious autoimmune diseases including rheumatic fever and post-infectious glomerulonephritis; solid cancers and tumor metastasis. 2-Cyclopentyl-6-methoxy-isonicotinic acid, which is also disclosed in WO201 1007324, is a useful intermediate for the synthesis of the pyridine-4-yl derivatives of formula (PD), wherein R^a is a cyclopentyl group.

In the process described in WO201 1007324, 2-cyclopentyl-6-methoxy-isonicotinic acid was prepared according to the following reaction scheme 1 :

Compound D Compound E

Rieke Zinc: cyclopentylzinc bromide;

PdCI₂(dppf)dcm: 1 ,1 ‘-Bis(diphenylphosphino)ferrocene-palladium(ll)dichloride

dichloromethane complex

However, the abovementioned process has drawbacks for larger scale, i.e. industrial scale synthesis of 2-cyclopentyl-6-methoxy-isonicotinic acid, for the following reasons:

a) The commercially available starting material, 2,6-dichloro-isonicotinic acid (Compound A) is expensive.

b) The conversion of Compound C to Compound D is cost-intensive. The reaction has to be performed under protective atmosphere with expensive palladium catalysts and highly reactive and expensive Rieke zinc complex. Such synthesis steps are expensive to scale up and it was therefore highly desired to find alternative synthesis methods.

Even though Goldsworthy, J. Chem. Soc. 1934, 377-378 discloses the preparation of 1 -cyclopentylethanone, which is a key building block in the new process of the present invention, by using ethyl 1 -acetoacetate as a starting material, this synthesis was far from being suitable in an industrial process. The reported yield was low (see also under “Referential Examples” below). Scheme 2

ethyl 1 -acetylcyclo- 1-cyclopentyl- pentanecarboxylate ethanone

Besides the early work by Goldsworthy there are several recent examples for the preparation of 1 -cyclopentylethanone described in the literature. Such examples include:

1 ) Addition of methyl lithium to a N-cyclopentanecarbonyl-N,0-dimethylhydroxylamine at -78°C in a yield of 77%. US2006/199853 A1 , 2006 and US2006/223884 A1 , 2006.

2) Addition of methyl lithium to a cyclopentyl carboxylic acid in diethylether at -78°C in a yield of 81 %. J. Am. Chem. Soc, 1983, 105, 4008-4017.

3) Addition of methylmagnesiumbromide to cyclopentanecarbonitrile.

Bull. Soc. Chim. Fr., 1967, 3722-3729.

4) Oxidation of 1 -cyclopentylethanol with chromtrioxide. US5001 140 A1 , 1991.

WO2009/71707 A1 , 2009.

5) Addition of cyclopentylmagnesium bromide to acetic anhydride at -78 °C with a yield of 54%. WO2004/74270 A2, 2004.

6) Synthesis of 1-cyclopentylethanone in 5 steps from cyclopentanone. Zhang, Pang; Li, Lian-chu, Synth. Commun., 1986, 16, 957-966.

However, the processes described in the above-listed publications are not efficient for scale-up since they require cryogenic temperatures, expensive starting materials, toxic reagents or many steps. The lack of an efficient process to manufacture 1 -cyclopentylethanone is further also mirrored by the difficulty in sourcing this compound on kilogram scale for a reasonable price and delivery time. Therefore, the purpose of the present invention is to provide a new, efficient and cost effective process for the preparation of 2-cyclopentyl-6-methoxy-isonicotinic acid, which is suitable for industrial scale synthesis.

Patent

https://patentscope.wipo.int/search/en/detail.jsf?docId=US133347630&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

Disclosed in WO2011007324, have immunomodulating activity through their S1P1/EDG1 receptor agonistic activity. Therefore, those pyridine-4-yl derivatives are useful for prevention and/or treatment of diseases or disorders associated with an activated immune system, including rejection of transplanted organs such as kidney, liver, heart, lung, pancreas, cornea, and skin; graft-versus-host diseases brought about by stem cell transplantation; autoimmune syndromes including rheumatoid arthritis, multiple sclerosis, inflammatory bowel diseases such as Crohn’s disease and ulcerative colitis, psoriasis, psoriatic arthritis, thyroiditis such as Hashimoto’s thyroiditis, uveo-retinitis; atopic diseases such as rhinitis, conjunctivitis, dermatitis; asthma; type I diabetes; post-infectious autoimmune diseases including rheumatic fever and post-infectious glomerulonephritis; solid cancers and tumor metastasis. 2-Cyclopentyl-6-methoxy-isonicotinic acid, which is also disclosed in WO2011007324, is a useful intermediate for the synthesis of the pyridine-4-yl derivatives of formula (PD), wherein R^a is a cyclopentyl group.

Rieke Zinc: cyclopentylzinc bromide;
PdCl₂(dppf)dcm: 1,1′-Bis(diphenylphosphino)ferrocene-palladium(II)dichloride dichloromethane complex

EXAMPLES

Example 1a

1-Cyclopentylethanone

Example 1 b

1-Cyclopentylethanone

Example 1c

1-Cyclopentylethanone

Example 1d

1-Cyclopentylethanone

Example 1e

1-Cyclopentylethanone

Example 1f

Tert-butyl 1-acetylcyclopentanecarboxylate

Example 1 g

Tert-butyl 1-acetylcyclopentanecarboxylate

Example 2

2-Cyclopentyl-6-hydroxyisonicotinic acid

Example 3

Methyl 2-cyclopentyl-6-hydroxyisonicotinate

Example 4a

Methyl 2-chloro-6-cyclopentylisonicotinate

Example 4b

Methyl 2-chloro-6-cyclopentylisonicotinate

Example 5

2-Cyclopentyl-6-methoxyisonicotinic acid

Example 6

Ethyl 4-cyclopentyl-2,4-dioxobutanoate

Example 7

Ethyl 3-cyano-6-cyclopentyl-2-hydroxyisonicotinate

REFERENTIAL EXAMPLES

Referential Example 1

Referential Example 2

PATENT

US8658675

https://www.google.com/patents/US8658675

Martin Bolli, Cyrille Lescop, Boris Mathys,Keith Morrison, Claus Mueller, Oliver Nayler,Beat Steiner,

novel compounds of Formula (I) that are agonists for the G protein-coupled receptor S1P1/EDG1 and have a powerful and long-lasting immunomodulating effect which is achieved by reducing the number of circulating and infiltrating T- and B-lymphocytes, without affecting their maturation, memory, or expansion. The reduction of circulating T-/B-lymphocytes as a result of S1P1/EDG1 agonism, possibly in combination with the observed improvement of endothelial cell layer function associated with S1P1/EDG1 activation, makes such compounds useful to treat uncontrolled inflammatory diseases and to improve vascular functionality. Prior art document WO 2008/029371 discloses compounds that act as S1P1/EDG1 receptor agonists and show an immunomodulating effect as described above. Unexpectedly, it has been found that the compounds of the present invention have a reduced potential to constrict airway tissue/vessels when compared to compounds of the prior art document WO 2008/029371. The compounds of the present invention therefore demonstrate superiority with respect to their safety profile, e.g. a lower risk of bronchoconstriction.

Examples of WO 2008/029371, which are considered closest prior art analogues are shown in FIG. 1.

The data on the constriction of rat trachea rings compiled in Table 1 illustrate the superiority of the compounds of the present invention as compared to compounds of prior art document WO 2008/029371.

For instance, the compounds of Example 1 and 6 of the present invention show a significantly reduced potential to constrict rat trachea rings when compared to the compounds of prior art Examples 222 and 226 of WO 2008/029371, respectively. Furthermore, the compounds of Example 1 and 6 of the present invention also show a reduced potential to constrict rat trachea rings when compared to the compounds of prior art Examples 196 and 204 of WO 2008/029371, respectively. These data demonstrate that compounds wherein R¹ represents 3-pentyl and R²represents methoxy are superior compared to the closest prior art compounds of WO 2008/029371, i.e. the compounds wherein R¹ represents an isobutyl and R²represents methoxy or wherein R¹ represents methyl and R² represents 3-pentyl. Moreover, also the compound of Example 16 of the present invention, wherein R¹is 3-methyl-but-1-yl and R² is methoxy, exhibits a markedly reduced potential to constrict rat trachea rings when compared to its closest analogue prior art Example 226 of WO 2008/029371 wherein R¹ is isobutyl and R² is methoxy.

The unexpected superiority of the compounds of the present invention is also evident from the observation that the compounds of Example 2 and 7 of the present invention show a markedly reduced potential to constrict rat trachea rings when compared to the compounds of prior art Examples 229 and 233 of WO 2008/029371, respectively. This proves that compounds wherein R¹ represents cyclopentyl and R² represents methoxy are superior compared to the closest prior art compounds of WO 2008/029371, i.e. the compounds wherein R¹represents methyl and R² represents cyclopentyl.

Preparation of Intermediates2-Chloro-6-methyl-isonicotinic acid

The title compound and its ethyl ester are commercially available.

2-(1-Ethyl-propyl)-6-methoxy-isonicotinic acid

a) To a solution of 2,6-dichloroisonicotinic acid (200 g, 1.04 mol) in methanol (3 L), 32% aq. NaOH (770 mL) is added. The stirred mixture becomes warm (34° C.) and is then heated to 70° C. for 4 h before it is cooled to rt. The mixture is neutralised by adding 32% aq. HCl (100 mL) and 25% aq. HCl (700 mL). The mixture is stirred at rt overnight. The white precipitate that forms is collected, washed with methanol and dried. The filtrate is evaporated and the residue is suspended in water (200 mL). The resulting mixture is heated to 60° C. Solid material is collected, washed with water and dried. The combined crops give 2-chloro-6-methoxy-isonicotinic acid (183 g) as a white solid; LC-MS: t_R=0.80 min, [M+1]⁺=187.93.

b) To a suspension of 2-chloro-6-methoxy-isonicotinic acid (244 g, 1.30 mol) in methanol (2.5 L), H₂SO₄ (20 mL) is added. The mixture is stirred at reflux for 24 h before it is cooled to 0° C. The solid material is collected, washed with methanol (200 mL) and water (500 mL) and dried under HV to give 2-chloro-6-methoxy-isonicotinic acid methyl ester (165 g) as a white solid; LC-MS: t_R=0.94 min, [M+1]⁺=201.89.

c) Under argon, Pd(dppf) (3.04 g, 4 mmol) is added to a solution of 2-chloro-6-methoxy-isonicotinic acid methyl ester (50 g, 0.248 mol) in THF (100 mL). A 0.5 M solution of 3-pentylzincbromide in THF (550 mL) is added via dropping funnel. Upon complete addition, the mixture is heated to 85° C. for 18 h before it is cooled to rt. Water (5 mL) is added and the mixture is concentrated. The crude product is purified by filtration over silica gel (350 g) using heptane:EA 7:3 to give 2-(1-ethyl-propyl)-6-methoxy-isonicotinic acid methyl ester (53 g) as a pale yellow oil; ¹H NMR (CDCl₃): δ0.79 (t, J=7.5 Hz, 6H), 1.63-1.81 (m, 4H), 2.47-2.56 (m, 1H), 3.94 (s, 3H), 3.96 (s, 3H), 7.12 (d, J=1.0 Hz, 1H), 7.23 (d, J=1.0 Hz, 1H).

d) A solution of 2-(1-ethyl-propyl)-6-methoxy-isonicotinic acid methyl ester (50 g, 0.211 mol) in ethanol (250 mL), water (50 mL) and 32% aq. NaOH (50 mL) is stirred at 80° C. for 1 h. The mixture is concentrated and the residue is dissolved in water (200 mL) and extracted with TBME. The org. phase is separated and washed once with water (200 mL). The TBME phase is discarded. The combined aq. phases are acidified by adding 25% aq. HCl and then extracted with EA (400+200 mL). The combined org. extracts are concentrated. Water (550 mL) is added to the remaining residue. The mixture is heated to 70° C., cooled to rt and the precipitate that forms is collected and dried to give the title compound (40.2 g) as a white solid; LC-MS: t_R=0.95 min, [M+1]⁺=224.04; ¹H NMR (D₆-DMSO): δ 0.73 (t, J=7.3 Hz, 6H), 1.59-1.72 (m, 4H), 2.52-2.58 (m, 1H), 3.88 (s, 3H), 7.00 (d, J=1.0 Hz, 1H), 7.20 (d, J=1.0 Hz, 1H).

2-Methoxy-6-(3-methyl-butyl)-isonicotinic acid

The title compound is prepared in analogy to 2-(1-ethyl-propyl)-6-methoxy-isonicotinic acid; LC-MS: t_R=0.94 min, [M+1]⁺=224.05; ¹H NMR (D₆-DMSO): δ 0.92 (d, J=5.8 Hz, 6H), 1.54-1.62 (m, 3H), 2.70-2.76 (m, 2H), 3.88 (s, 3H), 6.99 (s, 1H), 7.25 (s, 1H), 13.52 (s).

2-Cyclopentyl-6-methoxy-isonicotinic acid

The title compound is prepared in analogy to 2-(1-ethyl-propyl)-6-methoxy-isonicotinic acid; LC-MS: t_R=0.93 min, [M+1]⁺=222.02; ¹H NMR (CDCl₃): δ 1.68-1.77 (m, 2H), 1.81-1.90 (m, 4H), 2.03-2.12 (m, 2H), 3.15-3.25 (m, 1H), 3.99 (s, 3H), 7.18 (d, J=1.0 Hz, 1H), 7.35 (d, J=0.8 Hz, 1H).

2-Cyclohexyl-6-methoxy-isonicotinic acid

The title compound is prepared in analogy to 2-(1-ethyl-propyl)-6-methoxy-isonicotinic acid; LC-MS: t_R=0.98 min, [M+1]⁺=236.01; ¹H NMR (D₆-DMSO): δ 1.17-1.29 (m, 1H), 1.31-1.43 (m, 2H), 1.44-1.55 (m, 2H), 1.67-1.73 (m, 1H), 1.76-1.83 (m, 2H), 1.84-1.92 (m, 2H), 2.66 (tt, J=11.3, 3.3 Hz, 1H), 3.88 (s, 3H), 7.00 (d, J=1.0 Hz, 1H), 7.23 (d, J=1.0 Hz, 1H).

2-Cyclopentyl-N-hydroxy-6-methoxy-isonicotinamidine

a) A solution of 2-cyclopentyl-6-methoxy-isonicotinic acid methyl ester (3.19 g, 13.6 mmol) in 7 N NH₃ in methanol (50 mL) is stirred at 60° C. for 18 h. The solvent is removed in vacuo and the residue is dried under HV to give crude 2-cyclopentyl-6-methoxy-isonicotinamide (3.35 g) as a pale yellow solid; LC-MS**: t_R=0.57 min, [M+1]⁺=221.38.

b) Pyridine (8.86 g, 91.3 mmol) is added to a solution of 2-cyclopentyl-6-methoxy-isonicotinamide (3.35 g, 15.2 mmol) in DCM (100 mL). The mixture is cooled to 0° C. before trifluoroacetic acid anhydride (9.58 g, 45.6 mmol) is added portionwise. The mixture is stirred at 0° C. for 1 h before it is diluted with DCM (100 mL) and washed with sat. aq. NaHCO₃ solution (100 mL) and brine (100 mL). The separated org. phase is dried over MgSO₄, filtered and concentrated. The crude product is purified by CC on silica gel eluting with heptane:EA 9:1 to give 2-cyclopentyl-6-methoxy-isonicotinonitrile (2.09 g) as pale yellow oil; LC-MS**: t_R=0.80 min, [M+1]⁺=not detectable; ¹H NMR (D₆-DMSO): δ 1.61-1.82 (m, 6H), 1.94-2.03 (m, 2H), 3.16 (quint, J=7.8 Hz, 1H), 3.89 (s, 3H), 7.15 (s, 1H), 7.28 (s, 1H).

c) To a solution of 2-cyclopentyl-6-methoxy-isonicotinonitrile (2.09 g, 10.3 mmol) in methanol (100 mL), hydroxylamine hydrochloride (2.15 g, 31.0 mmol) and NaHCO₃ (3.04 g, 36.2 mmol) are added. The mixture is stirred at 60° C. for 18 h before it is filtered and the filtrate is concentrated. The residue is dissolved in EA (300 mL) and washed with water (30 mL). The washings are extracted back with EA (4×100 mL) and DCM (4×100 mL). The combined org. extracts are dried over MgSO₄, filtered, concentrated and dried under HV to give the title compound (2.74 g) as a white solid; LC-MS**: t_R=0.47 min, [M+1]⁺=236.24; ¹H NMR (D₆-DMSO): δ 1.61-1.82 (m, 6H), 1.92-2.01 (m, 2H), 3.04-3.13 (m, 1H), 3.84 (s, 3H), 5.90 (s, 2H), 6.86 (s, 1H), 7.13 (s, 1H), 9.91 (s, 1H).

2-Cyclopentyl-6-methoxy-isonicotinic acid hydrazide

a) To a solution of 2-cyclopentyl-6-methoxy-isonicotinic acid (2.00 g, 9.04 mmol), hydrazinecarboxylic acid benzyl ester (1.50 g, 9.04 mmol) and DIPEA (2.34 g, 18.1 mmol) in DCM (40 mL), TBTU (3.19 g, 9.94 mmol) is added. The mixture is stirred at rt for 2 h before it is diluted with EA (250 mL), washed twice with sat. aq. NaHCO₃ solution (150 mL) followed by brine (100 mL), dried over MgSO₄, filtered and concentrated. The crude product is purified by CC on silica gel eluting with heptane:EA 4:1 to give N′-(2-cyclopentyl-6-methoxy-pyridine-4-carbonyl)-hydrazinecarboxylic acid benzyl ester (2.74 g) as pale yellow oil; LC-MS**: t_R=0.74 min, [M+1]⁺=369.69; ¹H NMR (D₆-DMSO): δ 1.62-1.83 (m, 6H), 1.95-2.05 (m, 2H), 3.10-3.21 (m, 1H), 3.88 (s, 3H), 5.13 (s, 2H), 6.97 (s, 1H), 7.23 (s, 1H), 7.28-7.40 (m, 5H), 9.45 (s, 1H), 10.52 (s, 1H).

b) Pd/C (500 mg, 10% Pd) is added to a solution of N′-(2-cyclopentyl-6-methoxy-pyridine-4-carbonyl)-hydrazinecarboxylic acid benzyl ester (2.74 g, 7.42 mmol) in THF (50 mL) and methanol (50 mL). The mixture is stirred at rt under 1 bar of H₂ for 25 h. The catalyst is removed by filtration and the filtrate is concentrated and dried under HV to give the title compound (1.58 g) as an off-white solid; LC-MS**: t_R=0.51 min, [M+1]⁺=236.20; ¹H NMR (D₆-DMSO): δ 1.60-1.82 (m, 6H), 1.94-2.03 (m, 2H), 3.08-3.19 (m, 1H), 3.86 (s, 3H), 4.56 (s br, 2H), 6.93 (d, J=1.0 Hz, 1H), 7.20 (d, J=1.0 Hz, 1H), 9.94 (s, 1H).

3-Ethyl-4-hydroxy-5-methyl-benzonitrile

The title compound is prepared from 3-ethyl-4-hydroxy-5-methyl-benzaldehyde following literature procedures (A. K. Chakraborti, G. Kaur, Tetrahedron 55 (1999) 13265-13268); LC-MS: t_R=0.90 min; ¹H NMR (CDCl₃): δ1.24 (t, J=7.6 Hz, 3H), 2.26 (s, 3H), 2.63 (q, J=7.6 Hz, 2H), 5.19 (s, 1H), 7.30 (s, 2H).

3-Chloro-4-hydroxy-5-methyl-benzonitrile

The title compound is prepared from commercially available 2-chloro-6-methyl-phenol in analogy to literature procedures (see 3-ethyl-4-hydroxy-5-methyl-benzonitrile); LC-MS: t_R=0.85 min. ¹H NMR (CDCl₃): δ2.33 (s, 3H), 6.10 (s, 1H), 7.38 (s, 1H), 7.53 (d, J=1.8 Hz, 1H).

3-Ethyl-4,N-dihydroxy-5-methyl-benzamidine

The title compound is prepared from 3-ethyl-4-hydroxy-5-methyl-benzonitrile or from commercially available 2-ethyl-6-methyl-phenol following literature procedures (G. Trapani, A. Latrofa, M. Franco, C. Altomare, E. Sanna, M. Usala, G. Biggio, G. Liso, J. Med. Chem. 41 (1998) 1846-1854; A. K. Chakraborti, G. Kaur, Tetrahedron 55 (1999) 13265-13268; E. Meyer, A. C. Joussef, H. Gallardo, Synthesis 2003, 899-905); LC-MS: t_R=0.55 min; ¹H NMR (D₆-DMSO): δ 9.25 (s br, 1H), 7.21 (s, 2H), 5.56 (s, 2H), 2.55 (q, J=7.6 Hz, 2H), 2.15 (s, 3H), 1.10 (t, J=7.6 Hz, 3H).

3-Chloro-4,N-dihydroxy-5-methyl-benzamidine

The title compound is prepared from commercially available 2-chloro-6-methyl-phenol in analogy to literature procedures (e.g. B. Roth et al. J. Med. Chem. 31 (1988) 122-129; and literature cited for 3-ethyl-4,N-dihydroxy-5-methyl-benzamidine); 3-chloro-4-hydroxy-5-methyl-benzaldehyde: LC-MS: t_R=0.49 min, [M+1]⁺=201.00; ¹H NMR 82.24 (s, 2H), 2.35 (s, 4H), 5.98 (s br, 1H), 7.59 (d, J=1.8 Hz, 1H), 7.73 (d, J=1.8 Hz, 1H), 9.80 (s, 1H); 3-chloro-4,N-dihydroxy-5-methyl-benzamidine: ¹H NMR (D₆-DMSO): δ 2.21 (s, 3H), 5.72 (s br, 2H), 7.40 (s, 1H), 7.48 (s, 1H), 9.29 (s br, 1H), 9.48 (s br, 1H).

(R)-4-(2,2-Dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-N-hydroxy-5-methyl-benzamidine

a) To a solution of 3-ethyl-4-hydroxy-5-methyl-benzonitrile (2.89 g, 17.9 mmol) in THF (80 mL), (R)-(2,2-dimethyl-[1,3]dioxolan-4-yl)methanol (2.84 g, 21.5 mmol) followed by triphenylphosphine (5.81 g, 21.5 mmol) is added. The mixture is cooled with an ice-bath before DEAD (9.36 g, 21.5 mmol) is added dropwise. The mixture is stirred at rt for 1 h, the solvent is removed in vacuo and the residue is purified by CC on silica gel eluting with heptane:EA 85:15 to give (R)-4-(2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-5-methyl-benzonitrile (4.45 g) as a pale yellow oil; LC-MS**: t_R=0.75 min, [M+1]⁺=not detected; ¹H NMR (CDCl₃): δ1.25 (t, J=7.5 Hz, 3H), 1.44 (s, 3H), 1.49 (s, 3H), 2.34 (s, 3H), 2.65-2.77 (m, 2H), 3.80-3.90 (m, 2H), 3.94-4.00 (m, 1H), 4.21 (t, J=7.3 Hz, 1H), 4.52 (quint, J=5.8 Hz, 1H), 7.35 (s, 1H), 7.38 (s, 1H).

b) To a mixture of (R)-4-(2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-5-methyl-benzonitrile (4.45 g, 16.2 mmol) and NaHCO₃ (4.75 g, 56.6 mmol) in methanol (30 mL), hydroxylamine hydrochloride (3.37 g, 48.5 mmol) is added. The mixture is stirred at 60° C. for 18 h before it is filtered and the solvent of the filtrate is removed in vacuo. The residue is dissolved in EA and washed with a small amount of water and brine. The org. phase is separated, dried over MgSO₄, filtered, concentrated and dried to give the title compound (5.38 g) as a white solid; LC-MS**: t_R=0.46 min, [M+1]⁺=309.23; ¹H NMR (D₆-DMSO): δ 1.17 (t, J=7.5 Hz, 3H), 1.33 (s, 3H), 1.38 (s, 3H), 2.25 (s, 3H), 2.57-2.69 (m, 2H), 3.73-3.84 (m, 3H), 4.12 (t, J=7.0 Hz, 1H), 4.39-4.45 (m, 1H), 5.76 (s br, 2H), 7.34 (s, 1H), 7.36 (s, 1H), 9.47 (s, 1H).

(R)-3-Chloro-4-(2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-N-hydroxy-5-methyl-benzamidine

The title compound is obtained as a colorless oil (1.39 g) in analogy to (R)-4-(2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-N-hydroxy-5-methyl-benzamidine starting from 3-chloro-4-hydroxy-5-methyl-benzonitrile and L-α,β-isopropyliden glycerol; LC-MS: t_R=0.66 min, [M+H]⁺=314.96.

(S)-4-(3-Amino-2-hydroxypropoxy)-3-ethyl-5-methylbenzonitrile

a) To a solution of 3-ethyl-4-hydroxy-5-methyl-benzonitrile (5.06 g, 31.4 mmol) in THF (80 mL), PPh₃ (9.06 g, 34.5 mmol) and (R)-glycidol (2.29 mL, 34.5 mmol) are added. The mixture is cooled to 0° C. before DEAD in toluene (15.8 mL, 34.5 mmol) is added. The mixture is stirred for 18 h while warming up to rt. The solvent is evaporated and the crude product is purified by CC on silica gel eluting with heptane:EA 7:3 to give 3-ethyl-5-methyl-4-oxiranylmethoxy-benzonitrile (5.85 g) as a yellow oil; LC-MS: t_R=0.96 min; [M+42]⁺=259.08.

b) The above epoxide is dissolved in 7 N NH₃ in methanol (250 mL) and the solution is stirred at 65° C. for 18 h. The solvent is evaporated to give crude (S)-4-(3-amino-2-hydroxypropoxy)-3-ethyl-5-methylbenzonitrile (6.23 g) as a yellow oil; LC-MS: t_R=0.66 min; [M+1]⁺=235.11.

N—((S)-3-[2-Ethyl-4-(N-hydroxycarbamimidoyl)-6-methyl-phenoxy]-2-hydroxy-propyl)-2-hydroxy-acetamide

a) To a solution of (S)-4-(3-amino-2-hydroxypropoxy)-3-ethyl-5-methylbenzonitrile (6.23 g, 26.59 mmol) in THF (150 mL), glycolic acid (2.43 g, 31.9 mmol), HOBt (4.31 g, 31.9 mmol), and EDC hydrochloride (6.12 g, 31.9 mmol) are added. The mixture is stirred at rt for 18 h before it is diluted with sat. aq. NaHCO₃ and extracted twice with EA. The combined org. extracts are dried over MgSO₄, filtered and concentrated. The crude product is purified by CC with DCM containing 8% of methanol to give (S)—N-[3-(4-cyano-2-ethyl-6-methyl-phenoxy)-2-hydroxy-propyl]-2-hydroxy-acetamide (7.03 g) as a yellow oil; LC-MS: t_R=0.74 min, [M+1]⁺=293.10; ¹H NMR (CDCl₃): δ 1.25 (t, J=7.5 Hz, 3H), 2.32 (s, 3H), 2.69 (q, J=7.5 Hz, 2H), 3.48-3.56 (m, 3H), 3.70-3.90 (m, 3H), 4.19 (s, br, 3H), 7.06 (m, 1H), 7.36 (s, 1H), 7.38 (s, 1H).

b) The above nitrile is converted to the N-hydroxy-benzamidine according to literature procedures (e.g. E. Meyer, A. C. Joussef, H. Gallardo, Synthesis 2003, 899-905); LC-MS: t_R=0.51 min, [M+1]⁺=326.13; ¹H NMR (D₆-DMSO): δ 1.17 (t, J=7.4 Hz, 3H), 2.24 (s, 3H), 2.62 (q, J=7.4 Hz, 2H), 3.23 (m, 1H), 3.43 (m, 1H), 3.67 (m, 2H), 3.83 (s, 2H), 3.93 (m, 1H), 5.27 (s br, 1H), 5.58 (s br, 1H), 5.70 (s, 2H), 7.34 (s, 1H), 7.36 (s, 1H), 7.67 (m, 1H), 9.46 (s br, 1H).

(S)—N-(3-[2-Chloro-4-(N-hydroxycarbamimidoyl)-6-methyl-phenoxy]-2-hydroxy-propyl)-2-hydroxy-acetamide

The title compound is obtained as a beige wax (1.1 g) in analogy to N—((S)-3-[2-ethyl-4-(N-hydroxycarbamimidoyl)-6-methyl-phenoxy]-2-hydroxy-propyl)-2-hydroxy-acetamide starting from 3-chloro-4-hydroxy-5-methyl-benzonitrile; LC-MS: t_R=0.48 min, [M+H]⁺=331.94.

3-Chloro-N-hydroxy-4-methanesulfonylamino-5-methyl-benzamidine

a) A mixture of 4-amino-3-chloro-5-methylbenzonitrile (155 mg, 930 μmol) and methanesulfonylchloride (2.13 g, 18.6 mmol, 1.44 mL) is heated under microwave conditions to 150° C. for 7 h. The mixture is cooled to rt, diluted with water and extracted with EA. The org. extract is dried over MgSO₄, filtered and concentrated. The crude product is purified on prep. TLC using heptane:EA 1:1 to give N-(2-chloro-4-cyano-6-methyl-phenyl)-methanesulfonamide (105 mg) as an orange solid; LC-MS**: t_R=0.48 min; ¹H NMR (CDCl₃): δ2.59 (s, 3H), 3.18 (s, 3H), 6.27 (s, 1H), 7.55 (d, J=1.3 Hz, 1H), 7.65 (d, J=1.5 Hz, 1H).

b) Hydroxylamine hydrochloride (60 mg, 858 μmol) and NaHCO₃ (72 mg, 858 μmol) is added to a solution of N-(2-chloro-4-cyano-6-methyl-phenyl)-methanesulfonamide (105 mg, 429 μmol) in methanol (10 mL). The mixture is stirred at 65° C. for 18 h. The solvent is removed in vacuo and the residue is dissolved in a small volume of water (2 mL) and extracted three times with EA (15 mL). The combined org. extracts are dried over MgSO₄, filtered, concentrated and dried to give the title compound (118 mg) as a white solid; LC-MS**: t_R=0.19 min, [M+1]⁺=277.94; ¹H NMR (CDCl₃): δ2.57 (s, 3H), 3.13 (s, 3H), 6.21 (s, 1H), 7.49 (d, J=1.5 Hz, 1H), 7.63 (d, J=1.5 Hz).

3-Ethyl-N-hydroxy-4-methanesulfonylamino-5-methyl-benzamidine

a) In a 2.5 L three-necked round-bottom flask 2-ethyl-6-methyl aniline (250 g, 1.85 mol) is dissolved in DCM (900 mL) and cooled to 5-10° C. Bromine (310.3 g, 1.94 mol) is added over a period of 105 min such as to keep the temperature at 5-15° C. An aq. 32% NaOH solution (275 mL) is added over a period of 10 min to the greenish-grey suspension while keeping the temperature of the reaction mixture below 25° C. DCM (70 mL) and water (100 mL) are added and the phases are separated. The aq. phase is extracted with DCM (250 mL). The combined org. phases are washed with water (300 mL) and concentrated at 50° C. to afford the 4-bromo-2-ethyl-6-methyl-aniline (389 g) as a brown oil; ¹H NMR (CDCl₃): δ 1.27 (t, J=7.3 Hz, 3H), 2.18 (s, 3H), 2.51 (q, J=7.3 Hz, 2H), 3.61 (s br, 1H), 7.09 (s, 2H).

b) A double-jacketed 4 L-flask is charged with 4-bromo-2-ethyl-6-methyl-aniline (324 g, 1.51 mol), sodium cyanide (100.3 g, 1.97 mol), potassium iodide (50.2 g, 0.302 mol) and copper(I)iodide (28.7 g, 0.151 mol). The flask is evacuated three times and refilled with nitrogen. A solution of N,N′-dimethylethylenediamine (191.5 mL, 1.51 mol) in toluene (750 mL) is added. The mixture is heated to 118° C. and stirred at this temperature for 21 h. The mixture is cooled to 93° C. and water (1250 mL) is added to obtain a solution. Ethyl acetate (1250 mL) is added at 22-45° C. and the layers are separated. The org. phase is washed with 10% aq. citric acid (2×500 mL) and water (500 mL). The separated org. phase is evaporated to dryness to afford 4-amino-3-ethyl-5-methyl-benzonitrile (240 g) as a metallic black solid; ¹H NMR (CDCl₃): δ1.29 (t, J=7.5 Hz, 3H), 2.19 (s, 3H), 2.52 (q, J=7.3 Hz, 2H), 4.10 (s br, 1H), 7.25 (s, 2H).

c) The title compound is then prepared from the above 4-amino-3-ethyl-5-methyl-benzonitrile in analogy to 3-chloro-N-hydroxy-4-methanesulfonylamino-5-methyl-benzamidine; LC-MS**: t_R=0.26 min, [M+1]⁺=272.32.

3-Chloro-4-ethanesulfonylamino N-hydroxy-5-methyl-benzamidine

The title compound is prepared in analogy to 3-chloro-N-hydroxy-4-methanesulfonylamino-5-methyl-benzamidine using ethanesulfonylchloride; LC-MS**: t_R=0.27 min, [M+1]⁺=292.13; ¹H NMR (D₆-DMSO): δ 1.36 (t, J=7.5 Hz, 3H), 2.40 (s, 3H), 3.22 (q, J=7.5 Hz), 5.88 (s, 2H), 7.57 (d, J=1.5 Hz, 1H), 7.63 (d, J=1.5 Hz, 1H), 9.18 (s, 1H), 9.78 (s, 1H).

4-Benzyloxy-3-ethyl-5-methyl-benzoic acid

a) To a solution of 3-ethyl-4-hydroxy-5-methyl-benzaldehyde (34.9 g, 0.213 mol, prepared from 2-ethyl-6-methyl-phenol according to the literature cited for 3-ethyl-4,N-dihydroxy-5-methyl-benzamidine) in MeCN (350 mL), K₂CO₃ (58.7 g, 0.425 mol) and benzylbromide (36.4 g, 0.213 mol) are added. The mixture is stirred at 60° C. for 2 h before it is cooled to rt, diluted with water and extracted twice with EA. The org. extracts are washed with water and concentrated to give crude 4-benzyloxy-3-ethyl-5-methyl-benzaldehyde (45 g) as an orange oil. ¹H NMR (CDCl₃): δ1.29 (t, J=7.5 Hz, 3H), 2.40 (s, 3H), 2.77 (q, J=7.8 Hz, 2H), 4.90 (s, 2H), 7.31-7.52 (m, 5H), 7.62 (d, J=1.5 Hz, 1H), 7.66 (d, J=1.8 Hz, 1H), 9.94 (s, 1H).
b) To a mixture of 4-benzyloxy-3-ethyl-5-methyl-benzaldehyde (132 g, 0.519 mol) and 2-methyl-2-butene (364 g, 5.19 mol) in tert.-butanol (1500 mL), a solution of NaH₂PO₄ dihydrate (249 g, 2.08 mol) in water (1500 mL) is added. To this mixture, NaClO₂ (187.8 g, 2.08 mol) is added in portions. The temperature of the reaction mixture is kept below 30° C., and evolution of gas is observed. Upon completion of the addition, the orange bi-phasic mixture is stirred well for 3 h before it is diluted with TBME (1500 mL). The org. layer is separated and washed with 20% aq. NaHS solution (1500 mL) and water (500 mL). The org. phase is then extracted three times with 0.5 N aq. NaOH (1000 mL), the aq. phase is acidified with 25% aq. HCl (500 mL) and extracted twice with TBME (1000 mL). These org. extracts are combined and evaporated to dryness to give the title compound; ¹H NMR (D₆-DMSO): δ 1.17 (t, J=7.5 Hz, 3H), 2.31 (s, 3H), 2.67 (q, J=7.5 Hz, 2H), 4.86 (s, 2H), 7.34-7.53 (m, 5H), 7.68 (s, 2H), 12.70 (s, 1H).

Example 1 (S)-3-(2-Ethyl-4-{5-[2-(1-ethyl-propyl)-6-methoxy-pyridin-4-yl]-[1,2,4]oxadiazol-3-yl}-6-methyl-phenoxy)-propane-1,2-diol

a) To a solution of 2-(1-ethyl-propyl)-6-methoxy-isonicotinic acid (190 mg, 732 μmol) in THF (10 mL) and DMF (2 mL), DIPEA (190 mg, 1.46 mmol) followed by TBTU (235 mg, 732 μmol) is added. The mixture is stirred at rt for 10 min before (R)-4-(2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-N-hydroxy-5-methyl-benzamidine 226 mg, 732 μmol) is added. The mixture is stirred at rt for 1 h before it is diluted with EA and washed with water. The org. phase is separated and concentrated. The remaining residue is dissolved in dioxane (10 mL) and heated to 105° C. for 18 h. The mixture is cooled to rt, concentrated and the crude product is purified on prep. TLC plates using DCM containing 10% of methanol to give 4-{3-[4-((R)-2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-5-methyl-phenyl]-[1,2,4]oxadiazol-5-yl}-2-(1-ethyl-propyl)-6-methoxy-pyridine (256 mg) as a yellow oil; LC-MS: t_R=1.28 min, [M+H]⁺=496.23.

b) A solution of 4-{3-[4-((R)-2,2-dimethyl-[1,3]dioxolan-4-ylmethoxy)-3-ethyl-5-methyl-phenyl]-[1,2,4]oxadiazol-5-yl}-2-(1-ethyl-propyl)-6-methoxy-pyridine (250 mg, 504 μmol) in 4 M HCl in dioxane (10 mL) is stirred at rt for 90 min before it is concentrated. The crude product is purified on prep. TLC plates using DCM containing 10% of methanol to give the title compound (76 mg) as a pale brownish solid; LC-MS: t_R=1.12 min, [M+H]⁺=456.12; ¹H NMR (CDCl₃): δ0.85 (t, J=7.0 Hz, 6H), 1.33 (t, J=7.0 Hz, 3H), 1.70-1.89 (m, 4H), 2.42 (s, 3H), 2.61-2.71 (m, 1H), 2.78 (q, J=7.3 Hz, 2H), 3.82-4.00 (m, 4H), 4.04 (s, 3H), 4.14-4.21 (m, 1H), 7.34 (s, 1H), 7.46 (s, 1H), 7.86-7.91 (m, 2H).

Example 2 (S)-3-{4-[5-(2-Cyclopentyl-6-methoxy-pyridin-4-yl)-[1,2,4]oxadiazol-3-yl]-2-ethyl-6-methyl-phenoxy}-propane-1,2-diol

The title compound is prepared in analogy to Example 1 starting from 2-cyclopentyl-6-methoxy-isonicotinic acid; LC-MS: t_R=1.14 min, [M+H]⁺=454.16; ¹H NMR (CDCl₃): δ1.33 (t, J=7.5 Hz, 3H), 1.72-1.78 (m, 2H), 1.85-1.94 (m, 4H), 2.03-2.15 (m, 2H), 2.41 (s, 3H), 2.72 (d, J=5.3 Hz, 1H), 2.77 (q, J=7.5 Hz, 2H), 3.19-3.28 (m, 1H), 3.81-3.94 (m, 2 H), 3.95-3.98 (m, 2H), 4.02 (s, 3H), 4.14-4.21 (m, 1H), 7.31 (d, J=1.3 Hz, 1H), 7.51 (d, J=1.0 Hz, 1H), 7.88 (d, J=1.8 Hz), 7.89 (d, J=2.0 Hz, 1H).

PAPER

A practical synthesis of S1P receptor 1 agonist ACT-334441 (1) through late-stage convergent coupling of two key intermediates is described. The first intermediate is 2-cyclopentyl-6-methoxyisonicotinic acid whose skeleton was built from 1-cyclopentylethanone, ethyl oxalate, and cyanoacetate in a Guareschi–Thorpe reaction in 42% yield over five steps. The second, chiral intermediate, is a phenol ether derived from enantiomerically pure (R)-isopropylidene glycerol ((R)-solketal) and 3-ethyl-4-hydroxy-5-methylbenzonitrile in 71% yield in a one-pot reaction. The overall sequence entails 18 chemical steps with 10 isolated intermediates. All raw materials are cheap and readily available in bulk quantities, the reaction conditions match with standard pilot plant equipment, and the route reproducibly afforded 3–20 kg of 1 in excellent purity and yield for clinical studies.

Practical Synthesis of a S1P Receptor 1 Agonist via a Guareschi–Thorpe Reaction

RPL-554

• Molecular FormulaC₂₆H₃₁N₅O₄
• Average mass477.555

CFTR stimulator; PDE 3 inhibitor; PDE 4 inhibitor

RPL-554 is a mixed phosphodiesterase (PDE) III/IV inhibitor in phase II clinical development at Verona Pharma for the treatment of asthma, allergic rhinitis, chronic obstructive pulmonary disease (COPD) and inflammation.

RPL-554 is expected to have long duration of action and will be administered nasally thereby preventing gastrointestinal problems often resulting from orally administered PDE4 antiinflammatory drugs.

The company is now seeking licensing agreements or partnerships for the further development and commercialization of the drug.

RPL-554 (LS-193,855) is a drug candidate for respiratory diseases. It is an analog of trequinsin, and like trequinsin, is a dual inhibitor of the phosphodiesterase enzymes PDE-3 and PDE-4.^[1] As of October 2015, inhaled RPL-554 delivered via a nebulizer was in development for COPD and had been studied in asthma.^[2]

PDE3 inhibitors act as bronchodilators, while PDE4 inhibitors have an anti-inflammatory effect.^[1][3]

RPL554 was part of a family of compounds invented by Sir David Jack, former head of R&D for GlaxoSmithKline, and Alexander Oxford, a medicinal chemist; the patents on their work were assigned to Vernalis plc.^[4][5]:19-20

In 2005, Rhinopharma Ltd, acquired the rights to the intellectual property from Vernalis.^[5]:19-20 Rhinopharma was a startup founded in Vancouver, Canada in 2004 by Michael Walker, Clive Page, and David Saint, to discover and develop drugs for chronic respiratory diseases,^[5]:16 and intended to develop RPL-554, delivered with an inhaler, first for allergic rhinitis, then asthma, then forCOPD.^[5]:16-17 RPL554 was synthesized at Tocris, a contract research organization, under the supervision of Oxford, and was studied in collaboration with Page’s lab at King’s College, London.^[1] In 2006 Rhinopharma recapitalized and was renamed Verona Pharma plc.^[5]

This was first seen in April 2015 when it was published as a France national. Verona Pharma (formerly Rhinopharma), under license from Kings College via Vernalis, is developing the long-acting bronchodilator, RPL-554 the lead in a series dual inhibitor of multidrug resistant protein-4 and PDE 3 and 4 inhibiting trequinsin analogs which included RPL-565, for treating inflammatory respiratory diseases, such as allergic rhinitis, asthma, and COPD.

RPL554

Verona Pharma’s lead drug, RPL554, is a “first-in-class” inhaled drug under development for chronic obstructive pulmonary disease (COPD), asthma and cystic fibrosis. The drug is an inhibitor of the phosphodiesterase 3 (PDE3) and phosphodiesterase 4 (PDE4) enzymes, two enzymes known to be of importance in the development and progression of immunological respiratory diseases. The drug has the potential to act as both a bronchodilator and an anti-inflammatory which would significantly differentiate it from existing drugs.

RPL554 was selected from a class of compounds co-invented by Sir David Jack, the former Director of Research at Glaxo who led the team that discovered many of the commercially successful drugs in the respiratory market.

Verona Pharma has successfully completed two double-blind placebo controlled randomised Phase 2b studies of RPL554: one in mild to moderate asthma and another in mild to moderate COPD. The drug was found to be well tolerated, free from drug-related adverse effects (especially cardiovascular and gastro-intestinal effects) and generated significant bronchodilation.  Additionally, double-blind placebo controlled exploratory studies in healthy volunteers challenged with an inhaled irritant also generated consistent, clinically meaningful anti-inflammatory effects.

Verona Pharma is also carrying out exploratory studies to investigate the potential of RPL554 as a novel treatement for cystic fibrosis. In November 2014, the Company received a Venture and Innovation Award from the UK Cystic Fibrosis Trust to further such studies.

For further information on the potential of RPL554 for the treatment of respiratory diseases, refer to the peer-reviewed paper available on-line in the highly-respected medication journal, The Lancet Respiratory Medicine, entitled “Efficacy and safety of RPL554, a dual PDE3 and PDE4 inhibitor, in healthy volunteers and in patients with asthma or chronic obstructive pulmonary disease: findings from four clinical trials”.

The competitive advantages of RPL554 include the following:

• combining bronchodilator (PDE 3) and anti-inflammatory actions (PDE 4) in a single drug, something that is currently only achieved with a combination LABA and glucocorticosteroid inhaler,
• unique in not using steroids or beta agonists, which have known side effects,
• planned to be administered by nasal inhalation, thereby reducing the unwanted gastrointestinal side effects of many orally administered drugs.

History of Clinical Trials

• Following completion in May 2008 of toxicological studies of RPL554, the Company commenced in February 2009 a Phase I/IIa clinical trial of the drug at the Centre for Human Drug Research (CHDR) at Leiden in the Netherlands. In September 2009, the Company announced that it had successfully completed the trial, demonstrating that RPL554 has a good safety profile and has beneficial effects in terms of bronchodilation and bronchoprotection in asthmatics and a reduction in the numbers of inflammatory cells in the nasal passages of allergic rhinitis patients.
• In November 2010, the Company successfully completed a further trial that examined the safety and bronchodilator effectiveness of the drug administered at higher doses.
• In August 2011, the Company demonstrated that bronchodilation is maintained over a period of 6 days with daily dosing of RPL554 in asthmatics.
• In November 2011, the Company successfully demonstrated safety and bronchodilation of RPL554 in patients with mild to moderate forms of COPD.
• In March 2013, the Company demonstrated positive airway anti-inflammatory activity with respect to COPD at a clinical trial carried out at the Medicines Evaluation Unit (MEU) in Manchester, UK.

Synthesis

WO 2000058308

Cyclization of 1-(3,4-dimethoxyphenethyl)barbituric acid  in refluxing POCl3 produces the pyrimidoisoquinolinone , which is further condensed with 2,4,6-trimethylaniline  in boiling isopropanol to afford the trimethylphenylimino derivative . Subsequent alkylation of with N-(2-bromoethyl)phthalimide in the presence of K2CO3 and KI, followed by hydrazinolysis of the resulting phthalimidoethyl compound  yields the primary amine . This is finally converted into the title urea RPL 554 by reaction with sodium cyanate in aqueous HCl.

Example 1 : 9 Λ 0-Dimethoxy-2-(2.4-6-trimethy-phen yliminoY-3-(N-carbamoyl-2- aminoethylV3.4.6.7-tetrahydro-2H-pyrimido[6.1-a]isoquinolin-4-one

Sodium cyanate (6.0g, 0.092 mol) in water (100 ml) was added dropwise to a stirred solution of 9,10-Dimethoxy-2-(2,4,6-trimethylphenylimino)-3-(2-aminoethyl)-3,4,6,7- tetrahydro-2H-pyrimido[6,l-a]isoquinolin-4-one, prepared according to Preparation 4 above (20.0g, 0.046 mol) in water (600 ml) and IN ΗC1 (92 ml) at 80°C. After stirring for 2h at 80°C the mixture was cooled in an ice-bath and basified with 2N NaOH. The mixture was extracted with dichloromethane (3 x 200 ml) and the combined extract was dried (MgSO- ) and evaporated in vacuo. The resulting yellow foam was purified by column chromatography on silica gel eluting with CH2CI2 / MeOH (97:3) and triturated with ether to obtain the title compound as a yellow solid, 11.9g, 54%.

M.p.: 234-236°C m/z: C26H31N5O4 requires M=477 found (M+l) = 478

HPLC: Area (%) 99.50 Column ODS (150 x 4.6 mm)

MP pH3 KH₂PO₄ / CH3CN (60/40)

FR (ml/min) 1.0 RT (min) 9.25 Detection 250 nm

^lK NMR (300 MHz, CDCI3): δ 1.92 (1H, br s, NH), 2.06 (6H, s, 2xCH₃), 2.29 (3H, s, CH₃), 2.92 (2H, t, CH₂), 3.53 (2H, m, CH₂), 3.77 (3H, s, OCH3), 3.91 (3H, s, OCH3), 4.05 (2H, t, CH₂), 4.40 (2H, t, CH₂), 5.35 (2H, br s, NH₂), 5.45 (1H, s, C=CH), 6.68 (1H, s, ArH), 6.70 (1H, s, ArH), 6.89 (2H, s, 2xArH).

Preparation 1 : Synthesis of 2-Chloro-6.7-d-hydro-9.10-Dimethoxy-4H-pyrimido- [6,l-a]isoquinoHn-4-one (shown as (1) in Figure 1

A mixture of l-(3,4-dimethoxyphenyl) barbituric acid (70g, 0.24mol), prepared according to the method described in B. Lai et al. J.Med.Chem. 27 1470-1480 (1984), and phosphorus oxychloride (300ml, 3.22mol) was refluxed for 2.5h. The excess phosphorous oxychloride was removed by distillation (20mmHg) on wa ming. After cooling the residue was slurried in dioxan (100ml) and cautiously added to a vigorously stirred ice/water solution (11). Chloroform (11) was added and the resulting mixture was basified with 30% sodium hydroxide solution. The organic layer was separated and the aqueous phase further extracted with chloroform (2x750ml). The combined organic extracts were washed with water (1.51), dried over magnesium sulphate and concentrated in vacuo to leave a gummy material (90g). This was stirred in methanol for a few minutes, filtered and washed with methanol (200ml), diethyl ether (2x200ml) and dried in vacuo at 40°C to yield the title compound as a yellow/orange solid. 47g, 62%

(300MHz, CDCI3) 2.96(2H, t, C(₇) H₂); 3.96(6H, s, 2xOCH₃; 4.20(2H, t, C(₆₎ H₂); 6.61(1H, s, C₍₁₎ H); 6.76(1H, s, Ar-H); 7.10(1H, s, Ar-H). Preparation 2: 9.10-Dimethoxy-2-(2.4.6-trimethylphenyliminoV3.4.6.7- tetrahydro-2H-pyrimido[6.1-a]isoquinolin-4-one (shown as (2) in Figure 1

2-Chloro-9,10-dimethoxy-6,7-dihydro-4H-pyrimido[6,l-a]isoquinolin-4-one, prepared according to Preparation 1, (38.5g, 0.13 mol) and 2,4,6-trimethylaniline (52.7g, 0.39 mol) in propan-2-ol (3 1) was stirred and heated at reflux, under nitrogen, for 24h. After cooling to room temperature, the solution was evaporated in vacuo and the residue was purified by column chromatography on silica gel, eluting with CΗ2CI2 /

MeOH, initially 98:2, changing to 96:4 once the product began to elute from the column. The title compound was obtained with a slight impurity, (just above the product on tic). Yield 34.6g, 67%.

Preparation 3: 9.10-Dimethoxy-2-(2.4.6-trimethylphenyliminoV3-(2-N- phthalimidoethyπ-3.4.6.7-tetrahydro-2H-pyrimido[6.1-a]isoquinolin-4-one

(shown as (3 in Figure 1)

A mixture of 9,10-Dimethoxy-2-(2,4,6-trimethylphenylimino)-3,4,6,7-tetrahydro-2H- pyrimido[6,l-a]isoquinolin-4-one (which was prepared according to Preparation 2) (60.0g, 0.153 mol), potassium carbonate (191g, 1.38 mol), sodium iodide (137g, 0.92 mol) and N-(2-bromoethyl)phthalimide (234g, 0.92 mol) in 2-butanone (1500 ml) was stirred and heated at reflux, under nitrogen, for 4 days. After cooling to room temperature the mixture was filtered and the filtrate was evaporated in vacuo. The residue was treated with methanol (1000 ml) and the solid filtered off, washed with methanol and recrystallised from ethyl acetate to obtain the title compound as a pale yellow solid in yield 40. Og, 46%. Evaporation of the mother liquor and column chromatography of the residue on silica gel (CΗ2C-2 / MeOH 95:5) provided further product 11.7g, 13.5%. Preparation 4: 9.10-Dimethoxy-2-(2A6-trimethylphenylimino)-3-(2-arninoethyO- 3.4.6.7-tetrahydro-2H-pyrimido[6.1-a]isoquino-in-4-one (shown as (4) in Figure 1)

A mixture of 9,10-Dimethoxy-2-(2,4,6-trimethylphenylimino)-3-(2-N- phthalimidoethyl)-3,4,6,7-tetrahydro-2H-pyrimido[6,l-a]isoquinolin-4-one (22. Og, 0.039 mol), prepared according to Preparation 3, and hydrazine hydrate (11.3g, 0.195 mol) in chloroform (300 ml) and ethanol (460 ml) was stined at room temperature, under nitrogen, for 18h. Further hydrazine hydrate (2.9g, 0.05 mol) was added and the mixture was stirred a further 4h. After cooling in ice / water, the solid was removed by filtration and the filtrate evaporated in vacuo. The residue was dissolved in dichloromethane and the insoluble material was removed by filtration. The fitrate was dried (MgSO-i) and evaporated in vacuo to afford the title compound as a yellow foam in yield 16.2g, 96%.

PATENT

WO-2016128742

Novel crystalline acid addition salts forms of RPL-554 are claimed, wherein the salts, such as ethane- 1,2-disulfonic acid, ethanesulfonic acid, methanesulfonic acid, benzenesulfonic acid, p-toluenesulfonic acid, hydrochloric acid, hydrobromic acid, phosphoric acid or sulfuric acid. .

RPL554 (9, 10-dimethoxy-2-(2,4,6-trimethylphenylimino)-3-(/V-carbamoyl-2-aminoethyl)-3,4,6,7-tetrahydro-2H-pyrimido[6, l-a]isoquinolin-4-one) is a dual PDE3/PDE4 inhibitor and is described in WO 00/58308. As a combined PDE3/PDE4 inhibitor, RPL554 has both antiinflammatory and bronchodilatory activity and is useful in the treatment of respiratory disorders such as asthma and chronic obstructive pulmonary disease (COPD). The structure of RPL554 is shown below.

Owing to its applicability in the treatment of respiratory disorders, it is often preferable to administer RPL554 by inhalation. Franciosi et al. disclose a solution of RPL554 in a citrate-phosphate buffer at pH 3.2 (The Lancet: Respiratory Medicine 11/2013; l(9):714-27. DOI: 10.1016/S2213-2600(13)70187-5). The preparation of salts of RPL554 has not been described.

PATENT

http://www.google.ch/patents/WO2000058308A1?cl=en&hl=de

PATENT

http://www.google.ch/patents/WO2012020016A1?cl=en

U.S. Pat. No. 6,794,391, 7,378,424, and 7,105,663, which are each incorporated herein by reference, discloses compound RPL-554 (N-{2-[(2iT)-2-(mesityiimino)-9,10- dimethoxy-4-oxo-6,7-dihydro-2H-pyrimido[6,l-a]-isoquinolin-3 4H)-yl]ethyl}urea).

It would be beneficial to provide a composition of a stable polymorph of RPL-554, that has advanrtages over less stable polymorphs or amorphous forms, including

stability, compressibility, density, dissolution rates, increased potency or. lack toxicity.

References

1. Boswell-Smith V et al. The pharmacology of two novel long-acting phosphodiesterase 3/4 inhibitors, RPL554 [9,10-dimethoxy-2(2,4,6-trimethylphenylimino)-3-(n-carbamoyl-2-aminoethyl)-3,4,6,7-tetrahydro-2H-pyrimido[6,1-a]isoquinolin-4-one] and RPL565 [6,7-dihydro-2-(2,6-diisopropylphenoxy)-9,10-dimethoxy-4H-pyrimido[6,1-a]isoquinolin-4-one]. J Pharmacol Exp Ther. 2006 Aug;318(2):840-8. PMID 16682455
2.  Nick Paul Taylor for FierceBiotech. October 1, 2015 Verona sets sights on PhIIb after COPD drug comes through early trial
3.  Turner MJ et al. The dual phosphodiesterase 3 and 4 inhibitor RPL554 stimulates CFTR and ciliary beating in primary cultures of bronchial epithelia. Am J Physiol Lung Cell Mol Physiol. 2016 Jan 1;310(1):L59-70. PMID 26545902
4. Jump up^ see US20040171828, identified in the citations of PMID 16682455
5. ISIS Resources, PLC. August 23, 2006 Proposed Acquisition of Rhinopharma

REFERENCES

1: Calzetta L, Cazzola M, Page CP, Rogliani P, Facciolo F, Matera MG. Pharmacological characterization of the interaction between the dual phosphodiesterase (PDE) 3/4 inhibitor RPL554 and glycopyrronium on human isolated bronchi and small airways. Pulm Pharmacol Ther. 2015 Jun;32:15-23. doi: 10.1016/j.pupt.2015.03.007. Epub 2015 Apr 18. PubMed PMID: 25899618.

2: Franciosi LG, Diamant Z, Banner KH, Zuiker R, Morelli N, Kamerling IM, de Kam ML, Burggraaf J, Cohen AF, Cazzola M, Calzetta L, Singh D, Spina D, Walker MJ, Page CP. Efficacy and safety of RPL554, a dual PDE3 and PDE4 inhibitor, in healthy volunteers and in patients with asthma or chronic obstructive pulmonary disease: findings from four clinical trials. Lancet Respir Med. 2013 Nov;1(9):714-27. doi: 10.1016/S2213-2600(13)70187-5. Epub 2013 Oct 25. PubMed PMID: 24429275.

3: Wedzicha JA. Dual PDE 3/4 inhibition: a novel approach to airway disease? Lancet Respir Med. 2013 Nov;1(9):669-70. doi: 10.1016/S2213-2600(13)70211-X. Epub 2013 Oct 25. PubMed PMID: 24429260.

4: Calzetta L, Page CP, Spina D, Cazzola M, Rogliani P, Facciolo F, Matera MG. Effect of the mixed phosphodiesterase 3/4 inhibitor RPL554 on human isolated bronchial smooth muscle tone. J Pharmacol Exp Ther. 2013 Sep;346(3):414-23. doi: 10.1124/jpet.113.204644. Epub 2013 Jun 13. PubMed PMID: 23766543.

5: Gross N. The COPD pipeline XX. COPD. 2013 Feb;10(1):104-6. doi: 10.3109/15412555.2013.766103. PubMed PMID: 23413896.

6: Gross NJ. The COPD Pipeline XIV. COPD. 2012 Feb;9(1):81-3. doi: 10.3109/15412555.2012.646587. PubMed PMID: 22292600.

7: Boswell-Smith V, Spina D, Oxford AW, Comer MB, Seeds EA, Page CP. The pharmacology of two novel long-acting phosphodiesterase 3/4 inhibitors, RPL554 [9,10-dimethoxy-2(2,4,6-trimethylphenylimino)-3-(n-carbamoyl-2-aminoethyl)-3,4,6, 7-tetrahydro-2H-pyrimido[6,1-a]isoquinolin-4-one] and RPL565 [6,7-dihydro-2-(2,6-diisopropylphenoxy)-9,10-dimethoxy-4H-pyrimido[6,1-a]isoquino lin-4-one]. J Pharmacol Exp Ther. 2006 Aug;318(2):840-8. Epub 2006 May 8. PubMed PMID: 16682455.

RPL-554

Systematic (IUPAC) name
N-{2-[(2E)-2-(mesitylimino)-9,10-dimethoxy-4-oxo-6,7-dihydro-2H-pyrimido[6,1-a]-isoquinolin-3(4H)-yl]ethyl}urea
Identifiers
PubChem	CID 9934746
ChemSpider	8110374
Synonyms	9,10-Dimethoxy-2-(2,4,6-trimethylphenylimino)-3-(N-carbamoyl-2-aminoethyl)-3,4,6,7-tetrahydro-2H-pyrimido[6,1-a]isoquinolin-4-one
Chemical data
Formula	C26H31N5O4
Molar mass	477.554 g/mol

///////////RPL-554, LS-193,855, 298680-25-8, UNII:3E3D8T1GIX, RPL554, RPL 554, phase 2, Chronic Obstructive Pulmonary Diseases , COPD, Allergic Rhinitis, Asthma Therapy, Cystic Fibrosis, Inflammation, Bronchodilators

Cc3cc(C)cc(C)c3N=c2cc1-c(cc4OC)c(cc4OC)CCn1c(=O)n2CCNC(N)=O

Opicapone was approved by European Medicine Agency (EMA) on Jun 24, 2016. It was developed and marketed as Ongentys^® by Bial – Portela in EU.

Opicapone is a selective and reversible COMT inhibitor, used as adjunctive therapy for Parkinson’s disease.

Ongentys^® is available as hard capsules, containing 25 mg and 50 mg of opicapone. The recommended dose is 50 mg, taken once a day at bedtime, at least one hour before or after levodopa combination medicines.

 Chemical structures of tolcapone 1, entacapone 2, and nebicapone 3.

Opicapone

A preferred method of treatment of Parkinson’s disease is the administration of a combination of levodopa and a peripherally selective aromatic amino acid decarboxylase inhibitor (AADCI) together with a catechol-O-methyltransferase (COMT) inhibitor. The currently employed COMT inhibitors are tolcapone and entacapone. However, some authorities believe that each of these COMT inhibitors have residual problems relating to pharmacokinetic or pharmacodynamic properties, or to clinical efficiency or safety. Hence, not all patients get most benefit from their levodopa/AADCI/COMT inhibitor therapy.

Favoured new COMT inhibitors were disclosed in L. E. Kiss et al, J. Med. Chem., 2010, 53, 3396-3411 (D1), WO 2007/013830 (D2) and WO 2007/117165 (D3) which are believed to have particularly desirable properties so that patients can benefit from enhanced therapy.

D1, D2 and D3 also disclosed methods of preparing the new COMT inhibitors. Those processes, although effective, would benefit from an increase in yields. Other benefits which would be appropriate include those selected from reduction in number of process steps, reduction in number of unit operations, reduction of cycle-times, increased space yield, increased safety, easier to handle reagents/reactants and/or increase in purity of the COMT inhibitor, especially when manufacture of larger quantities are envisaged. A process has now been discovered that proceeds via a new intermediate which is suitable for manufacture of commercially useful quantities of a particularly apt COMT inhibitor in good yield. Additional benefits occur such as those selected from a reduced number of process steps and number of unit operations, reduced cycle-times, increased space yield, increased safety, with easier to handle reagents/reactants, improved impurity profile and/or good purity.

CLINICAL

https://clinicaltrials.gov/show/NCT01851850

NMR (DMSO-d6):
1H : 11.07 (2H, br, OH), 8.11 (1H, d, J = 2 Hz, H6), 7.73 (1H, d, J = 2 Hz, H2), 2.66 (3H, s, H15),2.24 (3H, s, H14).

13C : 175.2 (C7), 164.5 (C8), 150.4 (C12), 148.7 (C3), 146.3 (C4), 139.4 (C13), 137.8 (C5), 134.1(C10), 131.1 (C11), 122.7 (C9), 116.6 (C2), 115.7 (C6), 112.7 (C1), 17.9 (C14), 16.5 (C15).

Elemental Analysis:
(C15H10Cl2N4O6) C, H, N, S: Calc: C, 43.60; H, 2.44; N, 13.56; Found: C, 44; H, 2.3; N, 13.6.

PATENT

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2007013830&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

LEARMONTH, David Alexander; (PT).
KISS, Laszlo Erno; (PT).
LEAL PALMA, Pedro Nuno; (PT).
DOS SANTOS FERREIRA, Humberto; (PT).
ARAÚJO SOARES DA SILVA, Patrício Manuel Vieira; (PT)

PATENT

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2012107708

PATENT

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2007117165

scheme 1 depicts example how to produce a compound of the general formula IIB from a compound of the general formula IVB:

iii.

HB IVB
Scheme 1. Reagents: i. Piperidine, ethanol, reflux; ii. SO₂Cl₂, CCl₄, reflux; iii. POCI₃, 120 ⁰C, 18 h; iv. 50% H₂NOH, MeOH-H₂0, 1.25 mol % 1,10-phenanthroline hydrate.

The following reaction scheme 2 depicts an example how to produce certain compounds of general formula III:

I, R₈ = methyl III, R₈ = R₉ = H
iv.
R₉ = H

III, R₈ = R₉ = benzyl

Scheme 2. Reagents: i. 65 % HNO₃, AcOH; ii. 48 % HBr (aq), 140 ⁰C; iii. MeOH, HCl(g); iv. BnBr, K₂CO₃, CH₃CN, reflux; v. 3N NaOH, MeOH/H₂O.

The following reaction scheme 3 depicts an example how to produce the compound A, by activation of a compound according to general formula III followed by cyclisation involving condensation with a compound according to formula HB, dehydration and deprotection of the methyl residue protecting the hydroxyl group;

⁰C

compound A

REFERENCES

1: Bicker J, Alves G, Fortuna A, Soares-da-Silva P, Falcão A. A new PAMPA model using an in-house brain lipid extract for screening the blood-brain barrier permeability of drug candidates. Int J Pharm. 2016 Jan 30. pii: S0378-5173(16)30072-2. doi: 10.1016/j.ijpharm.2016.01.074. [Epub ahead of print] PubMed PMID: 26836708.

2: Devos D, Moreau C. Opicapone for motor fluctuations in Parkinson’s disease. Lancet Neurol. 2015 Dec 22. pii: S1474-4422(15)00346-4. doi: 10.1016/S1474-4422(15)00346-4. [Epub ahead of print] PubMed PMID: 26725545.

3: Ferreira JJ, Lees A, Rocha JF, Poewe W, Rascol O, Soares-da-Silva P; Bi-Park 1 investigators. Opicapone as an adjunct to levodopa in patients with Parkinson’s disease and end-of-dose motor fluctuations: a randomised, double-blind, controlled trial. Lancet Neurol. 2015 Dec 22. pii: S1474-4422(15)00336-1. doi: 10.1016/S1474-4422(15)00336-1. [Epub ahead of print] PubMed PMID: 26725544.

4: Rascol O, Perez-Lloret S, Ferreira JJ. New treatments for levodopa-induced motor complications. Mov Disord. 2015 Sep 15;30(11):1451-60. doi: 10.1002/mds.26362. Epub 2015 Aug 21. Review. PubMed PMID: 26293004.

5: Gonçalves D, Alves G, Fortuna A, Soares-da-Silva P, Falcão A. Development of a liquid chromatography assay for the determination of opicapone and BIA 9-1079 in rat matrices. Biomed Chromatogr. 2016 Mar;30(3):312-22. doi: 10.1002/bmc.3550. Epub 2015 Aug 17. PubMed PMID: 26147707.

6: Ferreira JJ, Rocha JF, Falcão A, Santos A, Pinto R, Nunes T, Soares-da-Silva P. Effect of opicapone on levodopa pharmacokinetics, catechol-O-methyltransferase activity and motor fluctuations in patients with Parkinson’s disease. Eur J Neurol. 2015 May;22(5):815-25, e56. doi: 10.1111/ene.12666. Epub 2015 Feb 4. PubMed PMID: 25649051.

7: Bonifácio MJ, Torrão L, Loureiro AI, Palma PN, Wright LC, Soares-da-Silva P. Pharmacological profile of opicapone, a third-generation nitrocatechol catechol-O-methyl transferase inhibitor, in the rat. Br J Pharmacol. 2015 Apr;172(7):1739-52. doi: 10.1111/bph.13020. Epub 2015 Jan 20. PubMed PMID: 25409768; PubMed Central PMCID: PMC4376453.

8: Kiss LE, Soares-da-Silva P. Medicinal chemistry of catechol O-methyltransferase (COMT) inhibitors and their therapeutic utility. J Med Chem. 2014 Nov 13;57(21):8692-717. doi: 10.1021/jm500572b. Epub 2014 Sep 2. PubMed PMID: 25080080.

9: Rocha JF, Falcão A, Santos A, Pinto R, Lopes N, Nunes T, Wright LC, Vaz-da-Silva M, Soares-da-Silva P. Effect of opicapone and entacapone upon levodopa pharmacokinetics during three daily levodopa administrations. Eur J Clin Pharmacol. 2014 Sep;70(9):1059-71. doi: 10.1007/s00228-014-1701-2. Epub 2014 Jun 14. PubMed PMID: 24925090.

10: Rocha JF, Santos A, Falcão A, Lopes N, Nunes T, Pinto R, Soares-da-Silva P. Effect of moderate liver impairment on the pharmacokinetics of opicapone. Eur J Clin Pharmacol. 2014 Mar;70(3):279-86. doi: 10.1007/s00228-013-1602-9. Epub 2013 Nov 24. PubMed PMID: 24271646.

11: Bonifácio MJ, Sutcliffe JS, Torrão L, Wright LC, Soares-da-Silva P. Brain and peripheral pharmacokinetics of levodopa in the cynomolgus monkey following administration of opicapone, a third generation nitrocatechol COMT inhibitor. Neuropharmacology. 2014 Feb;77:334-41. doi: 10.1016/j.neuropharm.2013.10.014. Epub 2013 Oct 19. PubMed PMID: 24148813.

12: Gonçalves D, Alves G, Fortuna A, Soares-da-Silva P, Falcão A. An HPLC-DAD method for the simultaneous quantification of opicapone (BIA 9-1067) and its active metabolite in human plasma. Analyst. 2013 Apr 21;138(8):2463-9. doi: 10.1039/c3an36671e. PubMed PMID: 23476919.

13: Rocha JF, Almeida L, Falcão A, Palma PN, Loureiro AI, Pinto R, Bonifácio MJ, Wright LC, Nunes T, Soares-da-Silva P. Opicapone: a short lived and very long acting novel catechol-O-methyltransferase inhibitor following multiple dose administration in healthy subjects. Br J Clin Pharmacol. 2013 Nov;76(5):763-75. doi: 10.1111/bcp.12081. PubMed PMID: 23336248; PubMed Central PMCID: PMC3853535.

14: Almeida L, Rocha JF, Falcão A, Palma PN, Loureiro AI, Pinto R, Bonifácio MJ, Wright LC, Nunes T, Soares-da-Silva P. Pharmacokinetics, pharmacodynamics and tolerability of opicapone, a novel catechol-O-methyltransferase inhibitor, in healthy subjects: prediction of slow enzyme-inhibitor complex dissociation of a short-living and very long-acting inhibitor. Clin Pharmacokinet. 2013 Feb;52(2):139-51. doi: 10.1007/s40262-012-0024-7. PubMed PMID: 23248072.

15: Palma PN, Bonifácio MJ, Loureiro AI, Soares-da-Silva P. Computation of the binding affinities of catechol-O-methyltransferase inhibitors: multisubstate relative free energy calculations. J Comput Chem. 2012 Apr 5;33(9):970-86. doi: 10.1002/jcc.22926. Epub 2012 Jan 25. PubMed PMID: 22278964.

16: Gonçalves D, Alves G, Soares-da-Silva P, Falcão A. Bioanalytical chromatographic methods for the determination of catechol-O-methyltransferase inhibitors in rodents and human samples: a review. Anal Chim Acta. 2012 Jan 13;710:17-32. doi: 10.1016/j.aca.2011.10.026. Epub 2011 Oct 20. Review. PubMed PMID: 22123108.

17: Kiss LE, Ferreira HS, Torrão L, Bonifácio MJ, Palma PN, Soares-da-Silva P, Learmonth DA. Discovery of a long-acting, peripherally selective inhibitor of catechol-O-methyltransferase. J Med Chem. 2010 Apr 22;53(8):3396-411. doi: 10.1021/jm1001524. PubMed PMID: 20334432.

////////BIA-9-1067,  ONO-2370,  BIA-91067, 923287-50-7, Opicapone, Catechol-O-methyl transferase, COMT inhibitor, Parkinson’s disease, PD, BIA 9-1067,  BIA 91067,  BIA-91067,  BIA91067, EU 2016

OC1=CC(C2=NC(C3=C(Cl)[N+]([O-])=C(C)C(Cl)=C3C)=NO2)=CC([N+]([O-])=O)=C1O

• Supporting Info

Naloxegol

Movantik; NKTR-118; NKTR118; UNII-44T7335BKE; NKTR 118

854601-70-0  cas

1354744-91-4 (Naloxegol Oxalate)

(4R,4aS,7S,7aR,12bS)-7-[2-[2-[2-[2-[2-[2-(2-methoxyethoxy)ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]ethoxy]-3-prop-2-enyl-1,2,4,5,6,7,7a,13-octahydro-4,12-methanobenzofuro[3,2-e]isoquinoline-4a,9-diol

Naloxegol oxalate (MovantikTM, Moventig)

Naloxegol (INN; PEGylated naloxol;^[1] trade names Movantik and Moventig) is a peripherally–selective opioid antagonistdeveloped by AstraZeneca, licensed from Nektar Therapeutics, for the treatment of opioid-induced constipation.^[2] It was approved in 2014 in adult patients with chronic, non-cancer pain.^[3] Doses of 25 mg were found safe and well tolerated for 52 weeks.^[4] When given concomitantly with opioid analgesics, naloxegol reduced constipation-related side effects, while maintaining comparable levels of analgesia.^[5]

Naloxegol Oxalate was approved by the U.S. Food and Drug Administration (FDA) on Sept 16, 2014, then approved by European Medicine Agency (EMA) on Dec 8, 2014. It was developed and marketed as Movantik^®(in the US)/Moventig^®(in EU) by AstraZeneca.

Naloxegol oxalate is an antagonist of opioid binding at the mu-opioid receptor. It is indicated for the treatment of opioid-induced constipation (OIC) in adult patients with chronic non-cancer pain.

Movantik^® is available as tablets for oral use, containing 12.5 mg or 25 mg of free Naloxegol. The recommended dose is 25 mg once daily (reduce to 12.5 mg if not tolerated).

Chemically, naloxegol is a pegylated (polyethylene glycol-modified) derivative of α-naloxol. Specifically, the 5-α-hydroxyl group of α-naloxol is connected via an ether linkage to the free hydroxyl group of a monomethoxy-terminated n=7 oligomer of PEG, shown extending at the lower left of the molecule image at right. The “n=7” defines the number of two-carbon ethylenes, and so the chain length, of the attached PEG chain, and the “monomethoxy” indicates that the terminal hydroxyl group of the PEG is “capped” with amethyl group.^[6] The pegylation of the 5-α-hydroxyl side chain of naloxol prevents the drug from crossing the blood-brain barrier(BBB).^[5] As such, it can be considered the antithesis of the peripherally-acting opiate loperamide which is utilized as an opiate-targeting anti-diarrheal agent that does not cause traditional opiate side-effects due to its inability to accumulate in the central nervous system in normal subjects.

Naloxegol was previously a Schedule II drug in the United States because of its chemical similarity to opium alkaloids, but was recently reclassified as a prescription drug after the FDA concluded that the impermeability of the blood-brain barrier to this compound made it non-habit-forming, and so without the potential for abuse — specifically, naloxegol was officially decontrolled on 23. January 2015. ^[7]

As an opiate antagonist, it is not expected to be capable of inducing the euphoria and anxiolytic effects which are generally cited as the desirable effects of commonly abused opiates (all of which are opiate agonists) if it were to cross the BBB; it would in fact reverse the effects of opiate drugs of abuse if it entered the central nervous system.

Naloxegol is an oral polyethylene glycol (PEG) derivative of naloxone, a peripherally acting µ-opioid receptor antagonist (PAMORA) with limited potential for interfering with centrally mediated opioid analgesia. The incorporation of a polyethylene glycol moiety aims at inhibiting naloxone’s capacity to cross the blood-brain barrier, while preserving the affinity for the µ-opioid receptor [1].

Opioid-induced bowel dysfunction (OIBD) represents a broad spectrum of symptoms that result from the actions of opioids on the CNS as well as the gastrointestinal tract. The majority of gastrointestinal effects seem to be mediated by the high number of µ-receptors that are expressed in the enteric nervous system. Naloxegol was more effective than placebo in increasing the number of spontaneous bowel movements in patients with opioid-induced constipation, including those with an inadequate response to laxatives.

Recognition of Naloxegol as a useful option in the treatment of opioid-induced constipation resulted in its approval by US-FDA for adult patients with chronic, non-cancer pain in 2014.
Naloxegol oxalate (XXI) is a peripherally acting l-opioid receptor antagonist that was approved in the USA and EU for the treatment of opioid-induced constipation in adults with chronic non-cancer pain. The drug, a pegylated version of naloxone, has significantly reduced central nervous system (CNS) penetration and works by inhibiting the binding of opioids in the gastrointestinal tract.152–154 Naloxegol oxalate was developed by Nektar and licensed to AstraZeneca. Although we were unable to find a single report in the primary or patent literature that describes the exact experimental procedures to prepare naloxegol oxalate, there havebeen reports on the preparation of closely related analogs155 with specific reports on improving the selectivity of the reduction step156 and the salt formation of the final drug substance.157 Taken together, the likely synthesis of naloxegol oxalate (XXI) is
described in Scheme 28. Naloxone (180) was treated with methoxyethyl chloride in the presence of Hunig’s base to give the protected ketone 181. Reduction of the ketone with potassium trisec-butylborohydride exclusively provided the a-alcohol 182 in 85% yield. Alternatively, sodium trialkylborohydrides could also be used to provide similar a-selective reduction in high yield.
Deprotonation of the alcohol with sodium hydride followed by alkylation with CH3(OCH2CH2)7Br (183) provided the pegylated intermediate 184 in 88% yield. Acidic removal of the methoxyethyl ether protecting group followed by treatment with oxalic acid and crystallization provided naloxegol oxalate (XXI) in good yield.

152. Corsetti, M.; Tack, J. Expert Opin. Pharmacol. 2015, 16, 399.
153. Garnock-Jones, K. P. Drugs 2015, 75, 419.
154. Leonard, J.; Baker, D. E. Ann. Pharmacother. 2015, 49, 360.
155. Bentley, M. D.; Viegas, T. X.; Goodin, R. R.; Cheng, L.; Zhao, X. US Patent
2005136031A1, 2005.
156. Cheng, L.; Bentley, M. D. WO Patent 2007124114A2, 2007.
157. Aaslund, B. L.; Aurell, C.-J.; Bohlin, M. H.; Sebhatu, T.; Ymen, B. I.; Healy, E. T.;
Jensen, D. R.; Jonaitis, D. T.; Parent, S. WO Patent 2012044243A1, 2012.
158. http://www.fda.gov/NewsEvents/Newsroom/PressAnnouncements/ucm4183

Naloxegol Synthesis

CREDIT

https://ayurajan.blogspot.in/2016/08/naloxegol.html

US20050136031A1: The patent reports detailed synthetic procedures to manufacture gram quantities of Naloxegol. The synthesis starts with Naloxone which was treated with methoxyethyl chloride in the presence of Hunig’s base to give the protected ketone. Reduction of the ketone with potassium tri-sec-butylborohydride exclusively provided the α-alcohol in 85% yield. Deprotonation of the alcohol with sodium hydride followed by alkylation with CH₃(OCH₂CH₂)₇Br  provided the pegylated Naloxone in 88% yield.

Identifications:

PATENT

US20060182692

PATENT

http://www.google.co.in/patents/WO2005058367A2?cl=en

EXAMPLE 4 SYNTHESIS OF PEG 3-NALθxoL [0211] The structure of the naloxol, an exemplary small molecule drug, is shown below.

Naloxol [0212] This molecule was prepared (having a protected hydroxyl group) as part of a larger synthetic scheme as described in Example 5.

EXAMPLE 5

[0213] α,β-PEGι-naloxol was prepared. The overview of the synthesis is provided below.

(3)

(4)

5.A. Synthesis of 3-MEM-naloxone

[0214] Diisopropylethylamine (390 mg, 3.0 mmole) was added to a solution of naloxone ^■ HCl ^• 2H₂O (200 mg, 0.50 mmole) in CH₂C1₂ (10 mL) with stining. Methoxyethyl chloride (“MEMCl,” 250 mg, 2.0 mmole) was then added dropwise to the above solution. The solution was stined at room temperature under N₂ overnight.

[0215] The crude product was analyzed by HPLC, which indicated that 3-

MEM-O-naloxone (1) was formed in 97% yield. Solvents were removed by rotary evaporation to yield a sticky oil.

5.B. Synthesis of α and β epimer mixture of 3-MEM-naloxoI (2)

[0216] 3 mL of 0.2 N NaOH was added to a solution of 3-MEM-naloxone

(1) (obtained from 5.A. above, and used without further purification) in 5mL of ethanol. To this was added a solution of NaBHLt (76 mg, 2.0 mmole) in water (1 mL) dropwise. The resulting solution was stined at room temperature for 5 hours. The ethanol was removed by rotary evaporation followed by addition of a solution of 0.1 N HCl solution to destroy excess NaBKj and adjust the pH to a value of 1. The solution was washed with CHC1₃ to remove excess methoxyethyl chloride and its derivatives (3 x 50 mL), followed by addition of K2OO₃ to raise the pH of the solution to 8.0. The product was then extracted with CHC1₃ (3 x 50 mL) and dried over Na₂SO₄. The solvent was removed by evaporation to yield a colorless sticky solid (192 mg, 0.46 mmole, 92% isolated yield based on naloxone ^• HCl ^• 2H2O).

[0217] HPLC indicated that the product was an α and β epimer mixture of

3-MEM-naloxol (2).

5.C. Synthesis of α and β epimer mixture of 6-CH₃-OCH₂CH₂-O-3-MEM- naloxol (3a).

[0218] NaH (60% in mineral oil, 55 mg, 1.38 mmole) was added into a solution of 6-hydroxyl-3-MEM-naloxol (2) (192 mg, 0.46 mmole) in dimethylformamide (“DMF,” 6 mL). The mixture was stined at room temperature under N₂ for 15 minutes, followed by addition of 2-bromoethyl methyl ether (320 mg, 2.30 mmole) in DMF (1 mL). The solution was then stirred at room temperature under N₂ for 3 hours.

[0219] HPLC analysis revealed formation of a mixture of α- and β-6-CH₃-OCH₂CH₂-0-3-MEM-naloxol (3) in about 88% yield. DMF was removed by a rotary evaporation to yield a sticky white solid. The product was used for subsequent transformation without further purification.

5.D. Synthesis of α and β epimer mixture of 6-CH₃-OCH₂CH₂-naloxoI (4)

[0220] Crude α- and β-6-CH₃-OCH₂CH₂-O-3-MEM-naloxol (3) was dissolved in 5 mL of CH₂C1₂ to form a cloudy solution, to which was added 5 mL of trifluoroacetic acid (“TFA”). The resultant solution was stined at room temperature for 4 hours. The reaction was determined to be complete based upon HPLC assay. CH₂C1₂ was removed by a rotary evaporator, followed by addition of 10 mL of water. To this solution was added sufficient K2OO₃ to destroy excess TFA and to adjust the pH to 8. The solution was then extracted with CHC1₃ (3 x 50 mL), and the extracts were combined and further extracted with 0.1 N HCl solution (3 x 50 mL). The pH of the recovered water phase was adjusted to a pH of 8 by addition of K₂CO_3>followed by further extraction with CHC1₃ (3 x 50 mL). The combined organic layer was then dried with Na2SO₄. The solvents were removed to yield a colorless sticky solid.

[0221] The solid was purified by passage two times through a silica gel column (2 cm x 30 cm) using CHCl₃/CH₃OH (30:1) as the eluent to yield a sticky solid. The purified product was determined by 1H NMR to be a mixture of α- and β epimers of 6-CH₃-OCH₂CH₂-naloxol (4) containing ca. 30% α epimer and ca. 70% β epimer [100 mg, 0.26 mmole, 56% isolated yield based on 6-hydroxyl-3-MEM- naloxol (2)].

[0222] 1H NMR (δ, ppm, CDC1₃): 6.50-6.73 (2 H, multiplet, aromatic proton of naloxol), 5.78 (1 H, multiplet, olefinic proton of naloxone), 5.17 (2 H, multiplet, olefinic protons of naloxol), 4.73 (1 H, doublet, C₅ proton of α naloxol), 4.57 (1 H, doublet, C₅ proton of β naloxol), 3.91 (1H, multiplet, C₆ proton of naloxol), 3.51-3.75 (4 H, multiplet, PEG), 3.39 (3 H, singlet, methoxy protons of PEG, α epimer), 3.36 (3 H, singlet, methoxy protons of PEG, β epimer), 3.23 (1 H, multiplet, C₆ proton of β naloxol), 1.46-3.22 (14 H, multiplet, protons of naloxol).

SYN 1

PATENT

https://www.google.com/patents/WO2012044243A1?cl=en

Naloxol-polyethylene glycol conjugates are provided herein in solid phosphate and oxalate salt forms. Methods of preparing the salt forms, and pharmaceutical compositions comprising the salt forms are also provided herein. ACKGROUND

Effective pain management therapy often calls for an opioid analgesic. In addition to the desired analgesic effect, however, certain undesirable side effects, such as bowel dysfunction, nausea, constipation, among others, can accompany the use of an opioid analgesic. Such side effects may be due to opioid receptors being present outside of the central nervous system, principally in the gastrointestinal tract. Clinical and preclinical studies support the use of mPEG₇-0-naloxol, a conjugate of the opioid antagonist naloxol and polyethylene glycol, to counteract undesirable side effects associated with use of opioid analgesics. When administered orally to a patient mPEG₇-0-naloxol largely does not cross the blood brain barrier into the central nervous system, and has minimal impact on opioid- induced analgesia. See, e.g., WO 2005/058367; WO 2008/057579; Webster et al., “NKTR-118 Significantly Reverses Opioid-Induced Constipation,” Poster 39, 20th AAPM Annual Clinical Meeting (Phoenix, AZ), October 10, 2009.

To move a drug candidate such as mPEG₇-O-naloxol to a viable pharmaceutical product, it is important to understand whether the drug candidate has polymorphic forms, as well as the relative stability and interconversions of these forms under conditions likely to be encountered upon large-scale production, transportation, storage and pre-usage preparation. Solid forms of a drug substance are often desired for their convenience in formulating a drug product. No solid form of mPEG₇-O-naloxol drug substance has been made available to date, which is currently manufactured and isolated as an oil in a free base form. Exactly how to accomplish this is often not obvious. For example the number of pharmaceutical products that are oxalate salts is limited. The free base form of mPEG₇-0-naloxol has not been observed to form a crystalline phase even when cooled to -60 °C and has been observed to exist as a glass with a transition temperature of

approximately -45 °C. Furthermore, mPEG₇-0-naloxol in its free base form can undergo oxidative degradation upon exposure to air. Care can be taken in handling the free base, for example, storing it under inert gas, to avoid its degradation. However, a solid form of mPEG₇-0-naloxol, preferably one that is stable when kept exposed to air, is desired

a naloxol-polyethlyene glycol conjugate oxalate salt, the salt comprising ionic species of mPEG₇-0-naloxol and oxalic acid. The formulas of mPEG₇-0-naloxol and oxalic acid are as follows:

In certain embodiments, the methods provided comprise dissolving mPEG₇-0- naloxol free base in ethanol; adding methyl t-butyl ether to the dissolved mPEG_?–

O-naloxol solution; adding oxalic acid in methyl t-butyl ether to the dissolved mPEG₇-0-naloxol over a period of at least 2 hours to produce a slurry; and filtering the slurry to yield the naloxol-polyethlyene glycol conjugate oxalate salt in solid form.

In certain embodiments, the methods provided comprise dissolving mPEG₇-0- naloxol free base in acetonitrile; adding water to the dissolved mPEG₇-0-naloxol solution; adding oxalic acid in ethyl acetate to the dissolved mPEG₇-0-naloxol over a period of at least 2 hours to produce a slurry; and filtering the slurry to yield the naloxol-polyethlyene glycol conjugate oxalate salt in solid form.

In some embodiments, the solid salt form of mPEG₇-0-naloxol is a crystalline form.

In certain embodiments a solid crystalline salt provided herein is substantially pure, having a purity of at least about 80%, at least about 85%, at least about 90%, at least about 92%, at least about 95%, at least about 96%, at least about 97%, at least about 98%, or at least about 99%.

In certain embodiments, the solid salt form of mPEG₇-0-naloxol is a phosphate salt.

In other embodiments, the solid mPEG₇-0-naloxol salt form is an oxalate salt. For instance, in some embodiments of solid oxalate salt forms provided herein, the solid mPEG₇-0-naloxol oxalate salt form is in Form A, as described herein. As another example, in some embodiments of solid oxalate salt forms provided herein, the solid mPEG₇-0-naloxol oxalate salt form is in Form B, as described herein. In yet other embodiments, an oxalate salt of mPEG₇-0-naloxol in solid form prepared according to the methods described herein is provided.

In yet other embodiments, an dihydrogenphosphate salt of mPEG₇-0-naloxol in solid form prepared according to the methods described herein is provided.

In certain embodiments of a solid mPEG₇-0-naloxol oxalate salt Form B provided herein, the salt form exhibits a single endothermic peak on differential scanning calorimetry between room temperature and about 150 °C. The single endothermic peak can occur, for instance, between about 91 °C to about 94 °C. For example, in some embodiments the endothermic peak is at about 92 °C, about 92.5 °C, or about93 °C.

PATENT

http://www.google.co.in/patents/WO2005058367A2?cl=en

PATENT

CN101033228A

PATENT

https://www.google.com/patents/CN102174049A?cl=en

References and notes

Naloxegol

Systematic (IUPAC) name
(5α,6α)-4,5-epoxy-6-(3,6,9,12,15,18,21-heptaoxadocos-1-yloxy)-17-(2-propen-1-yl)morphinan-3,14-diol
Clinical data
Trade names	Movantik, Moventig
AHFS/Drugs.com	movantik
License data	US FDA: Movantik
Pregnancy category	US: C (Risk not ruled out)
Routes of administration	Oral
Legal status
Legal status	US: ℞-only
Pharmacokinetic data
Protein binding	~4.2%
Metabolism	Hepatic (CYP3A)
Biological half-life	6–11 h
Excretion	Feces (68%), urine (16%)
Identifiers
CAS Number	854601-70-0
ATC code	A06AH03 (WHO)
PubChem	CID 56959087
ChemSpider	28651656
ChEBI	CHEBI:82975
Synonyms	NKTR-118
Chemical data
Formula	C34H53NO11
Molar mass	651.785 g/mol

//////////////Naloxegol, Movantik,  NKTR-118,  NKTR118,  UNII-44T7335BKE, NKTR 118, 854601-70-0, EU 2014, FDA 2014

COCCOCCOCCOCCOCCOCCOCCO[C@H]1CC[C@]2([C@H]3Cc4ccc(c5c4[C@]2([C@H]1O5)CCN3CC=C)O)O

CAS 128517-07-7

(1S,4S,7Z,10S,16E,21R)-7-ethylidene-4,21-di(propan-2-yl)-2-oxa-12,13-dithia-5,8,20,23-tetrazabicyclo[8.7.6]tricos-16-ene-3,6,9,19,22-pentone

(E)-N-(2-amino-4-fluorophenyl)-4-((3-(pyridin-3-yl)acrylamido)methyl)benzamide

Romidepsin, also known as Istodax, is an anticancer agent used in cutaneous T-cell lymphoma (CTCL) and other peripheral T-cell lymphomas (PTCLs). Romidepsin is a natural product obtained from the bacteria Chromobacterium violaceum, and works by blocking enzymes known as histone deacetylases, thus inducing apoptosis.^[1] It is sometimes referred to as depsipeptide, after the class of molecules to which it belongs. Romidepsin is branded and owned by Gloucester Pharmaceuticals, now a part of Celgene.^[2]

Romidepsin, a histone deacetylase inhibitor, originally developed by Fujisawa (now Astellas Pharma), causes cell cycle arrest,
differentiation, and apoptosis in various cancer cells.

In 2004, the FDA granted fast-track designation for romidepsin as monotherapy for the treatment of cutaneous T-cell lymphoma (CTCL) in patients who have relapsed following, or become refractory to, other systemic therapies. The FDA designated romidepsin as an orphan drug and it was approved in 2009 for this indication and it was commercialized in 2010. In 2007, another fast-track designation was granted for the product as monotherapy of previously treated peripheral T-cell lymphoma.

Romidepsin (FR901228) was originally discovered and isolated from the fermentation broth of Chromobacterium violaceum No. 968. It was identified through efforts in the search for novel agents which selectively reverse the morphological phenotype of Ras oncogene-transformed cells since the Ras signaling pathway plays a critical role in cancer development. Therefore, the drug could also have multiple molecular targets for its anticancer activity besides HDAC.

FR901228 is a bicyclic depsipeptide which is structurally unrelated to any known class of cyclic peptides with an unusual disulfide bond connecting a thiol and D-cysteine.

This drug is commercially produced by fermentation; however its interesting and novel structure warrants examination of its synthesis within the context of this review

Romidepsin is a histone deacetylase (HDAC) inhibitor.HDACs catalyze the removal of acetyl groups from acetylated lysine residues in histone and non-histone proteins, resulting in the modulation of gene expression.
Romidepsin is indicated for treatment of cutaneous T-cell lymphoma (CTCL) in patients who have received at least
one prior systemic therapy; treatment of peripheral T-cell lymphoma (PTCL) in patients who have received at least
one prior therapy.

Available as an injection, containing 10 mg of romidepsin and recommended dose is 14 mg/m2 administered intravenously over a 4-hour period on days 1, 8, and 15 of a 28-day cycle until disease progression or unacceptable toxicity.

History

Romidepsin was first reported in the scientific literature in 1994, by a team of researchers from Fujisawa Pharmaceutical Company (now Astellas Pharma) in Tsukuba, Japan, who isolated it in a culture of Chromobacterium violaceum from a soil sample obtained inYamagata Prefecture.^[3] It was found to have little to no antibacterial activity, but was potently cytotoxic against several human cancercell lines, with no effect on normal cells; studies on mice later found it to have antitumor activity in vivo as well.^[3]

The first total synthesis of romidepsin was accomplished by Harvard researchers and published in 1996.^[4] Its mechanism of actionwas elucidated in 1998, when researchers from Fujisawa and the University of Tokyo found it to be a histone deacetylase inhibitorwith effects similar to those of trichostatin A.^[5]

Clinical trials

Phase I studies of romidepsin, initially codenamed FK228 and FR901228, began in 1997.^[6] Phase II and phase III trials were conducted for a variety of indications. The most significant results were found in the treatment of cutaneous T-cell lymphoma (CTCL) and other peripheral T-cell lymphomas (PTCLs).^[6]

In 2004, romidepsin received Fast Track designation from the FDA for the treatment of cutaneous T-cell lymphoma, and orphan drugstatus from the FDA and the European Medicines Agency for the same indication.^[6] The FDA approved romidepsin for CTCL in November 2009^[7] and approved romidepsin for other peripheral T-cell lymphomas (PTCLs) in June 2011.^[8]

Mechanism of action

Romidepsin acts as a prodrug with the disulfide bond undergoing reduction within the cell to release a zinc-binding thiol.^[3][9][10] The thiol reversibly interacts with a zinc atom in the binding pocket of Zn-dependent histone deacetylase to block its activity. Thus it is anHDAC inhibitor. Many HDAC inhibitors are potential treatments for cancer through the ability to epigenetically restore normal expression of tumor suppressor genes, which may result in cell cycle arrest, differentiation, and apoptosis.^[11]

Adverse effects

The use of romidepsin is uniformly associated with adverse effects.^[12] In clinical trials, the most common were nausea and vomiting,fatigue, infection, loss of appetite, and blood disorders (including anemia, thrombocytopenia, and leukopenia). It has also been associated with infections, and with metabolic disturbances (such as abnormal electrolyte levels), skin reactions, altered taste perception, and changes in cardiac electrical conduction.^[12]

CLIP

http://www.scielo.br/scielo.php?script=sci_arttext&pid=S0103-50532012001200003

CLIP

Romidepsin was first isolated from the fermentation broth of Chromobacterium Violaceum WB968 in a nutrient
medium. Sterilized of 1% glucose and 1% bouillon solution were incubated with Chromobacterium Violaceum WB968, followed by further incubation with 1% glucose solution, 1% bouillon solution and adekanol gave the target romidepsin after extraction, silica gel chromatography and recrystallization.[Okuhara, M.; Goto, T.; Hori, Y. et al. US4977138A, 1990.]

The synthetic route was initiated by the deprotection L-(Fmoc)Thr-L-Val-OMe 1, subsequently coupled with
N-Alloc-S-Trt-D-Cys, followed by tosylation and then elimination to produce tripeptide 3 in the yield of 63.7% over four steps. The N-Alloc deprotection of 3 and then coupling with N-Fmoc-D-Valine were proceeded to provide tetrapeptide 4, which was subsequently removed Fmoc group to afford relative tetrapeptide 5 in 83.0% yield from compound 3. Condensation of 5 with β-hydroxy mercapto acid 6 was carried out by treating with benzotriazol-1-yl-oxy-tris-(dimethylamino)-phosphonium hexafluorphosphate (BOP) to give relative amide 7, and sequential hydrolysis yielded corresponding acid, which was performed by Mitsunobu macrolactonization
to produce depsipeptide 8 in 17.5% yield over three steps. Finally, romidepsin was obtained in the presence of iodine
in 81.0% yield and the overall yield of 7.5%.

The synthesis of intermediate β-hydroxy mercapto acid 6 commenced with the commercially available methyl 3,3-dimethoxypropionate 9. Nucleophilic addition of 9 with N,O-dimethylhydroxylamine provided Weinreb amide 10, followed by addition with lithium acetylide to give propargylic ketone 12 in the yield of 50.2% over two steps. Noyori’s asymmetric hydrogenation of ketone 12 provided (E)-alkene 14, which was removed the silyl group and then substituted with paratoluensulfonyl chloride to yield tosylate 15 in 40.6% yield across three steps. The dimethyl acetal of 15 was hydrolyzed to corresponding aldehyde by using lithium tetrafluoroborate,
which was immediately oxidized to relative carboxylic acid by applying Pinnick oxidation conditions. The trityl mercaptan was introduced by tosylate displacement to provide 6 in 65.0% yield over three steps and the overall yield of 13.3%.[2]

REF Greshock, T. J.; Johns, D. M.; Noguchi, Y., et al. Org. Lett. 2008, 10 (4), 613-616.

CLIP

Romidepsin (Istodax)
Romidepsin, a histone deacetylase inhibitor, originally developed by Fujisawa (now Astellas Pharma), causes cell cycle arrest,
differentiation, and apoptosis in various cancer cells.111 In 2004, the FDA granted fast-track designation for romidepsin as monotherapy for the treatment of cutaneous T-cell lymphoma (CTCL) in patients who have relapsed following, or become refractory
to, other systemic therapies. The FDA designated romidepsin as an orphan drug and it was approved in 2009 for this indication
and it was commercialized in 2010. In 2007, another fast-track designation was granted for the product as monotherapy of
previously treated peripheral T-cell lymphoma. Romidepsin (FR901228) was originally discovered and isolated from the fermentation
broth of Chromobacterium violaceum No. 968. It was identified through efforts in the search for novel agents which
selectively reverse the morphological phenotype of Ras oncogene-transformed cells since the Ras signaling pathway plays a
critical role in cancer development. Therefore, the drug could also have multiple molecular targets for its anticancer activity besides
HDAC.112 FR901228 is a bicyclic depsipeptide which is structurally unrelated to any known class of cyclic peptides with an unusual
disulfide bond connecting a thiol and D-cysteine. This drug is commercially produced by fermentation; however its interesting
and novel structure warrants examination of its synthesis within the context of this review.113,114 The synthesis of romidepsin
described is based on the total synthesis reported by the Williams115 and Simon groups (Scheme 20).116
L-Valine methyl ester (134) was coupled to N-Fmoc-L-threonine in the presence of the BOP reagent in 95% yield. The N-Fmoc protecting group was removed with Et2NH and the corresponding free amine was coupled to N-alloc-(S-triphenylmethyl)-D-cysteine with 1-ethyl-3-(3-dimethylaminopropyl)carbodiimide (EDCI) and HOBT in DMF and CH2Cl2 to yield the tripeptide 135 in good yield. The threonine residue of tripeptide 135 was then subjected to dehydrating conditions to give alkene 136 in 95% yield. The N-alloc protecting group of the dehydrated tripeptide 136 was removed with palladium and tin reagents and the corresponding free amine was subsequently coupled with N-Fmoc-D-valine to give tetrapeptide 137 in 83% yield. After removal of the N-Fmoc protecting group of compound 137 with Et2NH amine 138 was obtained in quantitative yield. The acid coupling partner 145 for
amine 138 was prepared as follows: methyl 3,3-dimethoxypropionate (139) was converted to its corresponding Weinreb amide by standard conditions and reacted with lithium acetylide 140 to give propargylic ketone 141 in 75% yield. Noyori’s asymmetric reduction of ketone 141 using ruthenium catalyst 142 gave the (R)-propargylic alcohol in 98% ee. This was followed by Red-Al reduction of the alkyne to selectively yield (E)-alkene 143 in 58% yield for the two steps. Liberation of the primary alcohol
with tetrabutylammonium fluoride (TBAF) followed by selective tosylation gave 144 in 70% yield in two steps. Hydrolysis of the dimethyl acetal of 144 with LiBF4 was followed by a Pinnick oxidation to give the corresponding carboxylic acid. The tosylate was displaced with trityl mercaptan in the presence of tert-butyl alcohol to give allylic alcohol 145 in 65% yield for the three steps.
Aminoamide 138 was then coupled to acid 145 using BOP to give peptide 146 in quantitative yield. The methyl ester of compound 146 was hydrolyzed with lithium hydroxide to provide the free carboxylic acid which underwent macrolactonization under Mitsunobu conditions in the presence of diisopropyl azodicarboxylate (DIAD) and triphenylphosine to give macrocycle 147 in 24% yield.
Finally, the disulfide linkage was formed by treating bis-tritylsulfane 147 with iodine in methanol at room temperature to give romidepsin (XIII) in 81% yield.

111 Bertino, E. M.; Otterson, G. A. Expert Opin. Invest. Drugs 2011, 20, 1151.
112. Furumai, R.; Matsuyama, A.; Kobashi, N.; Lee, K.-H.; Nishiyama, M.; Nakajima,
H.; Tanaka, A.; Komatsu, Y.; Nishino, N.; Yoshida, M.; Horinouchi, S. Cancer
Res. 2002, 62, 4916.
113. Verdine, G. L.; Vrolijk, N. H.; Bertel, S. WO 2008083288 A2, 2008.
114. Verdine, G. L.; Vrolijk, N. H. WO 2008083290 A1, 2008.
115. Greshock, T. J.; Johns, D. M.; Noguchi, Y.; Williams, R. M. Org. Lett. 2008, 10,
613.
116. Li, K. W.; Wu, J.; Xing, W.; Simon, J. A. J. Am. Chem. Soc. 1996, 118, 7237.

CLIP

http://pubs.rsc.org/en/content/articlelanding/2009/np/b817886k#!divAbstract

Williams’ improved synthesis of FK228.

Williams’ synthesis of the FK228 amide isostere (74).

References

External links

• Clinical trials of romidepsin at ClinicalTrials.gov

Romidepsin

Systematic (IUPAC) name
(1S,4S,7Z,10S,16E,21R)-7-ethylidene-4,21-diisopropyl-2-oxa-12,13-dithia-5,8,20,23-tetrazabicyclo[8.7.6]tricos-16-ene-3,6,9,19,22-pentone
Clinical data
Trade names	Istodax
MedlinePlus	a610005
License data	US FDA: Romidepsin
Pregnancy category	US: D (Evidence of risk)
Routes of administration	Intravenous infusion
Legal status
Legal status	US: ℞-only
Pharmacokinetic data
Bioavailability	Not applicable (IV only)
Protein binding	92–94%
Metabolism	Hepatic (mostly CYP3A4-mediated)
Biological half-life	3 hours
Identifiers
CAS Number	128517-07-7
ATC code	none
PubChem	CID 5352062
IUPHAR/BPS	7006
UNII	CX3T89XQBK
ChEBI	CHEBI:61080
ChEMBL	CHEMBL1213490
Synonyms	FK228; FR901228; Istodax
Chemical data
Formula	C24H36N4O6S2
Molar mass	540.695 g/mol

//////////fast-track designation, Romidepsin, Depsipeptide,  FK228,  Chromadax,  FR901228,  Istodax, FDA 2009, Fujisawa, Astellas Pharma, 128517-07-7

CC=C1C(=O)NC(C(=O)OC2CC(=O)NC(C(=O)NC(CSSCCC=C2)C(=O)N1)C(C)C)C(C)C

Bioprojet INNOVATOR

Jean-Charles Schwartz, Jeanne-Marie Lecomte

Pitolisant (INN) or tiprolisant (USAN) is a histamine receptor inverse agonist/antagonist selective for the H₃ subtype.^[1] It hasstimulant and nootropic effects in animal studies,^[2] and may have several medical applications, having been researched for the treatment of narcolepsy, for which it has been granted orphan drug status in the EU and US.^[3][4] It is currently in clinical trials forschizophrenia and Parkinson’s disease.^[4][5][6]

Pitolisant hydrochloride was approved by European Medicine Agency (EMA) on Mar 31, 2016. It was developed and marketed as Wakix^® by Bioprojet in EU.

Pitolisant hydrochloride is an antagonist/inverse agonist of the histamine H3 receptor, which is indicated in adults for the treatment of narcolepsy with or without cataplexy.

Wakix^® is available as tablet for oral use, containing 4.5 mg and 18 mg of Pitolisant hydrochloride. The initial dose of 9 mg (two 4.5 mg, tablets) per day, and it should be used at the lowest effective dose, depending on individual patient response and tolerance, according to an up-titration scheme, without exceeding the dose of 36 mg/day.

Pitolisant was developed by Jean-Charles Schwartz, Walter Schunack and colleagues after the former discovered H₃ receptors.^[7]Pitolisant was the first clinically used H₃ receptor inverse agonist.

Pitolisant, also known as Tiprolisant, is a histamine receptor inverse agonist/antagonist selective for the H3 subtype. It has stimulant and nootropic effects in animal studies, and may have several medical applications, having been researched for the treatment of narcolepsy, for which it has been granted orphan drug status in the EU and US. It is currently in clinical trials for schizophrenia and Parkinson’s disease. Pitolisant was the first clinically used H3 receptor inverse agonist.

The European Medicines Agency (EMA) has recommended granting marketing authorization for pitolisant (Wakix, Bioprojet Pharma) for narcolepsy with or without cataplexy, the agency announced today.

Narcolepsy is a rare sleep disorder that affects the brain’s ability to regulate the normal sleep-wake cycle, leading to excessive daytime sleepiness, including the sudden urge to sleep, and disturbed night-time sleep. Some patients also experience sudden episodes of cataplexy, potentially causing dangerous falls and increasing the risks for accidents, including car accidents. Symptoms of narcolepsy can be severe and significantly reduce quality of life.

Pitolisant “will add to the available treatment options for narcolepsy. It is a first-in-class medicine that acts on histamine H3 receptors in the brain. This leads to increased histamine release in the brain, thereby enhancing wakefulness and alertness,” the EMA notes in a news release.

The EMA recommendation for approval of pitolisant is based on an evaluation of all available safety and efficacy data conducted by the Committee for Medicinal Products for Human Use (CHMP). The data include two pivotal placebo-controlled trials involving 259 patients, as well as one uncontrolled, open-label study involving 102 patients with narcolepsy and one supportive study in 105 patients.

The studies showed that pitolisant was effective in reducing excessive daytime sleepiness in patients with narcolepsy. The beneficial effect of the drug on cataplexy was demonstrated in one of the pivotal studies as well as in the supportive study.

No major safety concerns with pitolisant emerged in testing. Insomnia, headache, and nausea were among the most common adverse effects observed in the clinical trials, and the CHMP decided on measures to mitigate these risks, the EMA said. The CHMP also requested the company conduct a long-term safety study to further investigate the safety of the drug when used over long periods.

Pitolisant for narcolepsy received orphan designation from the Committee for Orphan Medicinal Products in 2007. Orphan designation provides medicine developers access to incentives, such as fee reductions for scientific advice, with the aim of encouraging the development of treatments for rare disorders.

The CHMP opinion will now be sent to the European Commission for the adoption of a decision on a European Union–wide marketing authorization. Once that has been granted, each member state will decide on price and reimbursement based on the potential role/use of this medicine in the context of its national health system.

Narcolepsy-cataplexy.

Narcolepsy-cataplexy, or Gelineau syndrome, is a rare but serious disorder characterized by excessive daytime sleepiness which can be an extreme hindrance to normal professional and social activities, and which is accompanied by more or less frequent attacks of cataplexy (a sudden loss of muscle tone triggered by emotions as varied as laughter or fear) and erratic episodes of REM sleep (during wakefulness and during sleep), sometimes associated with hypnagogic hallucinations. Moreover, individuals with narcolepsy have various degrees of cognitive impairment and tend to be obese (reviewed by Dauvilliers et al., Clin. Neurophysiol., 2003, 114, 2000; Baumann and Bassetti, Sleep Med. Rev., 2005, 9, 253).

The disorder is caused by the loss of a group of neurons in the brain which produce two peptides, orexins, also known as hypocretins, located in the anterior hypothalamus and projecting to the main groups of aminergic neurons which regulate wakefulness and sleep. Patients with the disorder generally have very low levels of orexins in cerebrospinal fluid. Orexin knock-out mice display many of the symptoms seen in narcoleptic subjects, confirming the role of these peptides and thereby providing an excellent animal model of the disease (Chemelli et al., Cell, 1999, 98, 437).

Several types of treatments which can improve the symptoms of narcolepsy already exist, although they do not completely relieve symptoms and, furthermore, can cause significant side effects limiting their usefulness.

For instance, amphetamines or analogues such as methylphenidate which release catecholamines are used to treated daytime sleepiness, but these agents induce a state of excessive excitation as well as cardiovascular disturbances and also carry a potential for drug addiction.

Modafinil, a drug whose mechanism of action is unclear, also improves daytime sleepiness without causing as many side effects as amphetamines. Nonetheless, its efficacy is limited and it can cause headaches and nausea, particularly at high doses. Moreover amphetamines and/or modafinil do not appear to improve some of the most disabling symptoms of the disease, particularly cataplexy attacks, cognitive deficits and weight gain. With regard to cataplexy, treatments include antidepressants and oxybate. Effectiveness of the former has not been demonstrated (Cochrane Database Syst. Rev., 2005, 20, 3), and the latter is a drug of illegal abuse and its use is restricted.

It has also been shown that histamine H3 receptor antagonists induce the activation of histaminergic neurons in the brain which release histamine, a neurotransmitter with a crucial role in maintaining wakefulness (Schwartz et al., Physiol. Rev. 1991, 71, 1).

PATENT

https://patentscope.wipo.int/search/en/detail.jsf?docId=WO2006084833&recNum=1&maxRec=&office=&prevFilter=&sortOption=&queryString=&tab=PCTDescription

Pharmaceutical products with histamine H3 receptor ligand properties and 0 subsequent pharmacological activities thereof are described in EP-980300. An especially important product among those disclosed is 1-[3-[3-(4- chlorophenyl)propoxy] propyl]-piperidine. This compound is disclosed as the free base and as the oxalate salt.

5 The use of 1-[3-[3-(4-chlorophenyl)propoxy]propyl]-piperidine as the free base is limited because of its oily nature. On the contrary, 1-[3-[3-(4- chlorophenyl)propoxy]propyl]-piperidine oxalate is a crystalline substance but its low aqueous solubility (0.025 g/ml at 230C) also limits its use as a
pharmaceutical ingredient.
0
Subsequent patents EP-1100503 and EP-1428820 mention certain salts of 1- [3-[3-(4-chlorophenyl)propoxy]propyl]-piperidine. However, the only one specifically described is the oxalate salt. The crystalline monohydrochloride salt is not described.

Example 1 : 1-[3-[3-(4-chlorophenyl)propoxy]propyl]-piperidine

According to the method disclosed in EP-982300, Example 78, sodium 3-piperidinopropanolate (2.127 kg; 12.88 mol), 3-(4-chlorophenyl)propyl mesylate (1.121 kg; 4.51 mol) and 0.322 mol of 15-crown-5 in 4.5 kg of dry toluene were refluxed for 4 hours. The solvent was evaporated and the residue purified by column chromatography on silica gel (eluent: methylene chloride/methanol (90/10)). The obtained oil was distilled in a fractionating equipment at reduced pressure (0.3-0.7 mmHg) and with a heating jacket at 207-210⁰C. The head fractions and the distilled fraction at 0.001-0.010 mmHg with a jacket temperature of 180-200⁰C were collected. The obtained oil (1.0 kg; 3.38 mol) corresponds to 1-[3-[3-(4-chlorophenyl)propoxy] propyl]-piperidine. Yield 75%.

Example 2: 1-[3-[3-(4-chlorophenyl)propoxy]propyl]-piperidine
monohydrochloride

Preparation

Distilled 1-[3-[3-(4-chlorophenyl)propoxy]propyl]-piperidine (1.0 kg) and anhydrous ethyl acetate (4.5 kg) are transferred to a 10-L glass vessel fitted with a cooling bath and a gas inlet. A stream of gaseous hydrogen chloride is bubbled in the reaction mixture at 20-25⁰C.

The pH of the solution is checked by taking a 0.5 mL sample of the reaction mixture and diluting it with 5 mL of deionized water. The final pH must be about 3-4.

The mixture is cooled to -10°C-(-12°C) and stirred at this temperature for 1 h. The precipitate is filtered by using a sintered glass filter and washed with 0.5 L of anhydrous ethyl acetate previously cooled to 0-5⁰C. The product is dried in a vacuum oven at 5O⁰C for a minimum period of 12 hours. The resulting crude 1 -[3-[3-(4-chlorophenyl)propoxy]propyl]-piperidine monohydrochloride weighs 1.10 kg.

Purification

A mixture of the above-described crude, 3.98 kg of anhydrous ethyl acetate and 0.35 kg of /-propanol is heated slowly at 55-6O⁰C in a 10-L glass vessel fitted with a heating and cooling system. When the solution has been completed, it is filtered through a heat-isolated sintered glass filter, keeping the temperature at 55-6O⁰C. The solution is transferred to a 10 L glass vessel and the mass is slowly cooled to 0-5⁰C for about 1 hour. The mixture is stirred at this temperature for 1 hour and the precipitate is filtered through a sintered glass filter. The solid is washed with a mixture of 1.6 kg of anhydrous ethyl acetate and 0.14 kg of /-propanol cooled at 0-5⁰C. The solid is dried in a vacuum oven at 5O⁰C for a minimum period of 12 hours. M. p. 117-119⁰C. Yield 80%.
IR spectrum (KBr): bands at 1112 and 1101 (C-O Ether/ St. asym), 2936 and 2868 (Alkane CH(CH₂)) / St.), 1455 (Alkane CH(CH₂)) / Deform.), 2647 and 2551 (Amine Salt / St.), 1492 (Amine / St.), 802 (Aromatic / Deform.) cm^“1.

SEE

Eur. J. Pharm. Sci. 2001, 13, 249–259.

US2004220225A1.

CN101155793A

CN101171009A

References

Pitolisant

Names
IUPAC name 1-{3-[3-(4-Chlorophenyl)propoxy]propyl}piperidine
Other names BF2.649
Identifiers
CAS Number	903576-44-3
ChEMBL	ChEMBL462605
ChemSpider	8123714
Jmol 3D model	Interactive image
PubChem	9948102
InChI[show]
SMILES[show]
Properties
Chemical formula	C17H26ClNO
Molar mass	295.846 g/mol
Pharmacology
ATC code	N07XX11 (WHO)

//////////Pitolisant Hydrochloride, Wakix, histamine H3 receptor antagonist/inverse agonist, narcolepsy, orphan drug, tiprolisant

ClC1=CC=C(CCCOCCCN2CCCCC2)C=C1

Fortune India has published list of 500 mid-size companies who ranked them on various parameters based on the results of 2013-2014. Ajanta pharma features very prominently in the lists. Ajanta’s ranking on various pararmeters is given below:

Ranked 3’d largest Wealth Creator on 5 year CAGR 93.14%
Ranked 10th on Capital Employed (ROCE)
Ranked 21st in Net profit .
Ranked 182nd in Sales
On 17’th August 2015, Fortune India organized an award function to present the awards to Top 10 largest weatth creator companies and Ajanta is one of those elite companies.

The awards were presented by Mr. Piyush Goyal, Minister of State-Power,Coal& New and Renewable Energy, Govt. of India to Mr. yogesh Agrawal, Managing Director and Mr.Rajesh Agrawal,Jt.Managing Director of the company.

Ajanta Pharma, “One of the Giants of Tomorrow” – Fortune India

We are pleased to share with you that Ajanta Pharma has been honoured as “ONE OF THE GIANTS OF TOMORROW” by prestigious Fortune India magazine on 19^th August 2016 at New Delhi.

The honour was conferred to our Managing Director, Mr. Yogesh Agrawal and our Jt. Managing Director, Mr. Rajesh Agrawal at the hands of Hon. Mr. Nitin Gadkari, Union Minister for Road Transport and Highways and Shipping, Govt of India.  This is the 2^nd year in row where Ajanta has received the recognition from Fortune India.

Rajesh Agrawal (left), Ajanta Pharma’s joint managing director, with brother Yogesh, who is also managing director of the company, at their Kandivli facility

Ajanta Pharma needed a shot of its own medicine, an energiser like 30-Plus. It found its antidote in the new generation of Agrawals: Mannalal’s sons, Yogesh and Rajesh.

“When I joined Ajanta (in 2000), and realised what was going on, I wanted to run away. I thought to myself, ‘Why did I return from the US? I could have had a job there,’” says Rajesh, 39, Ajanta’s joint managing director, who has a management degree from Bentley College, Massachusetts. “It was tough in the beginning, especially the situation with creditors and debtors.”

Together, Rajesh and his older brother Yogesh, 43, who is managing director, changed Ajanta’s trajectory by focusing on the ‘specialty’ generic drug market and putting an end to the company’s legacy businesses, which included OTC drug sales and supplying drugs to government health agencies in India and other countries.

This was a risky move, but it has paid off. Ajanta Pharma closed FY15 with a consolidated net sales of Rs 1,481 crore and a net profit of Rs 310 crore (this is a compound annual growth rate, or CAGR, of 57 percent for four years since 2011). In terms of net sales, it recorded a CAGR of 31 percent for the same period. This growth has come on a low base, but the signs are encouraging. Its market value currently stands at around Rs 13,500 crore; this is a 65-fold growth in 15 years.

Read more: http://forbesindia.com/article/super-50-companies-2015/ajanta-pharma-the-small-big-dream/40691/1#ixzz4Igtudt24

References

http://www.ajantapharma.com/%5CAdminData%5CNewsRelease%5Ca8ea3740-99fc-4d81-be39-741f6ea95c542015-FortuneIndiapresentsawardtoAjantaPharma.pdf

https://www.linkedin.com/company/263285

/////////Ajanta Pharma, “One of the Giants of Tomorrow” ,  Fortune India, AWARD, Fortune India, RAJESH AGRAWAL

CAS .146510-36-3(Olanexidine free form), 

Imidodicarbonimidic diamide, N-((3,4-dichlorophenyl)methyl)-N’-octyl

C17H27Cl2N5
Formula Weight:	372.341

CAS 799787-53-4(Olanexidine Gluconate)

568.49
Formula	C17H27Cl2N5 ● C6H12O7

1-(3,4-Dichlorobenzyl)-5-octylbiguanide mono-D-gluconate

オラネキシジングルコン酸塩 Olanexidine Gluconate C17H27Cl2N5▪C6H12O7 : 568.49 [799787-53-4]

Indication：Bacterial infection

Otsuka (Originator)

• Marketed Bacterial infections

Most Recent Events

• 16 Sep 2015 Launched for Bacterial infections (Prevention) in Japan (Topical)
• 03 Jul 2015 Registered for Bacterial infections (Prevention) in Japan (Topical) – First global approval
• 30 Sep 2014 Preregistration for Bacterial infections (Prevention) in Japan (Topical)

SEE ALSO

Olanexidine hydrochloride [USAN]

146509-94-6 HCL
RN: 218282-71-4 HCL HYDRATE
UNII: R296398ALN

Molecular Formula, C17-H27-Cl2-N5.Cl-H.1/2H2-O

Molecular Weight, 835.6192

Imidodicarbonimidic diamide, N-((3,4-dichlorophenyl)methyl)-N’-octyl-, monohydrochloride, hydrate (2:1)

INTRODUCTION

Olanexidine gluconate was approved by Pharmaceuticals and Medical Devices Agency of Japan (PMDA) on Jul 03, 2015. It was developed and marketed as Olanedine^® by Otsuka in Japan.

Olanexidine gluconate is an antiseptic/disinfectant compound with potent bactericidal activity against Gram-negative and Gram-positive bacteria, for use in preparing patients for surgery and preventing of postoperative bacterial infections.

Olanedine^® is available as topical solution (1.5%), containing 3 g/200 mL, 0.15 g/10 mL and 0.375 g/25 mL, and the recommendation is applying appropriate amount of the drug.

PRODUCT PATENT

WO 2004105745

Kazuyoshi Miyata, Yasuhide Inoue, Akifumi Hagi, Motoya Kikuchi, Hitoshi Ohno, Kinji Hashimoto, Kinue Ohguro, Tetsuya Sato,Hidetsugu Tsubouchi, Hiroshi Ishikawa,Takashi Okamura, Koushi Iwata,

SYNTHESIS

PATENT

CN1065453A

http://www.google.com.au/patents/CN1065453A?cl=en

PATENT

WO2008026757A1

https://google.com/patents/WO2008026757A1?cl=en

Example 1: l-cyano-3-n-octylguanidine

A 7.00-kg quantity of Compound (4) (54.16 mol) was dissolved in 105 liters of ethyl acetate, and the resulting mixture was cooled to 5°C or below. A 2.66-kg quantity of concentrated sulfuric acid (27.12 mol) was added thereto dropwise at a temperature of 4O⁰C or below while stirring. To the thus- obtained suspension of 1/2 sulfate of Compound (4) was added 5.06 kg of sodium dicyanamide (56.83 mol), and the resulting suspension was heated under reflux for 7 hours. The reaction solution was cooled to 40⁰C or below, and 70 liters of water was added thereto. Subsequently, the resulting solution was heated to 80 to 90⁰C (internal temperature) to distill the ethyl acetate off. The remaining liquid was cooled to 40⁰C or below, and 70 liters of toluene was then added thereto, followed by the extraction of 1-cyano — 3-n-octyl guanidine at about 50⁰C. The extracted toluene layer was washed with 35 liters of water at about 50⁰C and cooled to 10⁰C or below, followed by stirring for about 30 minutes. The resulting precipitated crystals were separated and washed with 7 liters of toluene. The resulting crystals were dried at 40⁰C for 7.5 hours, yielding l-cyano-3-n- octylguanidine. 2007/067107

-16-

Yield: 9.11 kg (The yield was 85.7% based on the Compound(4).) White crystals having a melting point of 69 to 74⁰C (no clear melting point was observed)

IR(KBr) spectrum: 3439, 3296, 2916, 2164, 1659, 1556, 1160, 718, and 572 cm^“1

Thermogravimetric measurement/differential thermal analysis: 73.5°C (weak), an endothermic peak at 77.5⁰C

¹H-NMR(CDCl₃) spectrum: 0.88 ppm (t, J = 6.6 Hz, 3H), 1.20-1.38 ppm (m, 10H), 1.43-1.62 ppm (m, 2H), 3.17 ppm (dd, J = 6.9 Hz, J = 6.0 Hz, 2H), 5.60-5.70 ppm (bs, 2H), 5.80-5.95 ppm (bs, IH)

Reference Example 2: Acidolysis of 1- (3,4-dichlorobenzyl) -5- octylbiguanide dihydrochloride

A 1-g quantity of 1- (3, 4-dichlorobenzyl) -5-octyl biguanide dihydrochloride was dissolved in 15 ml of 10% ethanol, followed by refluxing for 5 hours. HPLC analysis was conducted under the conditions described below.

The yield of 1-[N- (3,4-dichlorobenzyl) carbamoyl-3- octyl]guanidine (holding time: 9.84 minutes) was 0.91%, and the yield of 1- (N-octyl-carbamoyl) -3- (3, 4-dichlorobenzyl) guanidine

(holding time: 10.54 minutes) was 0.22%.

HPLC analysis conditions:

Column: YMC AM302 4.6 mm I. D. x 150 mm

Eluate: MeCN/0.05 M aqueous solution of sodium 1- octanesulfonate/acetic acid = 700/300/1

Detector: UV 254 nm

The physical property values of the resulting 1-[N- (3,4- dichlorobenzyl) carbamoyl-3-octyl] guanidine were as follows: NMR (DMSO-de) δ: 0.86 (3H, t, J = 6.0 Hz), 1.07-1.35 (1OH, m) , 1.35-1.49 (2H, m) , 2.95-3.15 (2H, m) , 4.12 (2H, d, J = 6.3 Hz), 6.78-7.40 (4H, m) , 7.23 (IH, dd, J = 2.1 Hz, J = 8.4 Hz), 7.46 (IH, d, J = 2.1 Hz), 7.54 (IH, d, J = 8.4 Hz)

The physical property values of the resulting 1- (N-octyl- carbamoyl) -3- (3, 4-dichlorobenzyl) guanidine were as follows: NMR (DMSO-d₆) δ: 0.85 (3H, t, J = 6.6 Hz), 1.02-1.40 (12H, m) , 2.89-2.95 (2H, m) , 4.33 (2H, bs) , 5.76-7.00 (4H, m) , 7.28 (IH, dd, J = 2.1 Hz, J = 8.1 Hz), 7.52 (IH, d, J = 2.1 Hz), 7.58 (IH, d, J = 8.1 Hz)

Example 1: 1- (3, 4-dichlorobenzyl) -5-octylbiguanide monohydrochloride 1/2 hydrate

A 9.82-g quantity of Compound (2) (0.05 mol) and 10.63 g of 3, 4-dichlorobenzylamine (0.05 mol) were added to 49 ml of butyl acetate, followed by refluxing for 6 hours. The reaction solution was concentrated under reduced pressure, and a mixture of 12 ml of water and 47 ml of isopropyl alcohol was added and dissolved into the remainder. To the thus-obtained solution was added, dropwise, 10.13 g of concentrated hydrochloric acid. The resulting mixture was stirred at 28 to 30⁰C for 30 minutes, and the precipitated crystals were then filtered out. The thus- obtained crystals were washed with a small amount of isopropyl alcohol, yielding 23.42 g of (non-dried) 1- (3, 4-dichlorobenzyl) – 5-octylbiguanide dihydrochloride. The resulting crystals were suspended in 167 ml of water without drying, the suspension was then stirred at 25 to 27°C for 2 hours, followed by separation of the crystals by filtration. The thus-obtained crystals were washed with a small amount of water and dried at 40⁰C for 20 hours, yielding 17.05 g of 1- (3, 4-dichlorobenzyl) -5-octyl biguanide monohydrochloride 1/2 hydrate having a purity of 99.9% at a yield of 81.6%.

Example 2 : 1- (3, 4-dichlorobenzyl) -5-octylbiguanide dihydrochloride

A 100-g quantity of Compound (4) (0.774 mol) was dissolved in 1 liter of n-butyl acetate, and 37.6 g of concentrated sulfuric acid (0.383 mol) was added thereto while stirring. To the thus-obtained suspension of 1/2 sulfate of Compound (4) was added 68.9 g of sodium dicyanamide (0.774 mol), 7107

-18- and the resulting suspension was heated under reflux for 3 hours. The reaction solution was cooled to about 20⁰C, and the organic layer thereof was sequentially washed with about 500 ml each of (i) 5% hydrochloric acid, (ii) 5% aqueous caustic soda solution, (iii) 5% aqueous sodium bicarbonate solution, and (iv) water.

To the thus-obtained n-butyl acetate solution of Compound (2) were added 118.5 g of Compound (3) (0.673 mol) and then 58.4 ml of concentrated hydrochloric acid while stirring. The reaction solution was heated, and about 800 ml of n-butyl acetate was distilled off under atmospheric pressure (ordinary pressure) , followed by heating the reaction solution under reflux for 3.5 hours . Subsequently, the reaction solution was cooled to about 40⁰C, and 900 ml of isopropanol, 100 ml of water, and 134 ml of concentrated hydrochloric acid were added thereto. The mixture was stirred at 60 to 70°C for 1 hour and cooled to 10⁰C or below and the precipitated crystals were then separated. The resulting crystals were washed with 200 ml of isopropanol and dried at 6O⁰C, yielding 1- (3, 4-dichlorobenzyl) -5-octylbiguanide dihydrochloride. Yield: 243.8 g (The yield was 81.3% based on the Compound (3).) Melting point: 228.9⁰C IR(KBr) spectrum: 2920, 1682, 1634, 1337, 1035, 820, and 640 cm^“1

PATENT

WO2004105745A1

PATENT

WO2009142715A1

PATENT

https://www.google.com/patents/US8334248

Olanexidine is a compound with high bactericidal activity having the chemical name 1-(3,4-dichlorobenzyl)-5-octylbiguanide. Research has been carried out into bactericides containing, olanexidine hydrochloride as an active ingredient (see Japanese Patent No. 2662343, etc.).

Olanexidine has very poor solubility in water, and hitherto known salts of olanexidine are also poorly soluble in water. For example, the solubility at 0° C. of olanexidine hydrochloride in water has been measured to be less than 0.05% (W/V), and the solubility of free olanexidine is a further order of magnitude less than this. Consequently, sufficient bactericidal activity cannot be expected of an aqueous solution merely having olanexidine dissolved therein, and moreover, depending on the conditions the olanexidine may precipitate out.

In the case of making an aqueous preparation of olanexidine in particular, to make the concentration of the olanexidine sufficient for exhibiting effective bactericidal activity, and to reduce the possibility of the olanexidine precipitating out, it has thus been considered necessary to use a dissolution aid such as a surfactant.

EXAMPLE 1 Preparation of an Aqueous Solution Aqueous Solution 1

20.9 g (50 mmol) of olanexidine hydrochloride hemihydrate was added to 250 mL of a 1 N aqueous sodium hydroxide solution, and the suspension was stirred for 1.5 hours at room temperature (25° C.). The solid was filtered off, and washed with water. The solid obtained was further suspended in 250 mL of purified water, the suspension was stirred for 5 minutes at room temperature, and the solid was filtered off, and washed with water. This operation was carried out once more to remove sodium chloride formed. The solid obtained (free olanexidine) was put into purified water in which 8.9 g (50 mmol) of gluconolactone had been dissolved, and the mixture was stirred at room temperature until the solid dissolved, and then purified water was further added to give a total volume of 300 mL. The concentration of olanexidine in the aqueous solution obtained was measured by using high performance liquid chromatography to be 6% in terms of free olanexidine.

This aqueous solution was still transparent and colorless even after being left for several months at room temperature.

CLIP

http://dmd.aspetjournals.org/content/28/12/1417/F9.expansion.html

REFERENCES

http://www.otsukakj.jp/en/news/photo/photo-14423716650.pdf

//////////Olanexidine Gluconate, OPB-2045G, (Olanedine^®, Approved, japan 2015-07-03, Olanedine,  Otsuka, PMDA, Olanexidine, オラネキシジングルコン酸塩 , Gluconate olanexidin,  Olanedine,  OPB-2045,  OPB 2045G, JAPAN 2015

CCCCCCCCN=C(N)NC(=NCC1=CC(=C(C=C1)Cl)Cl)N

Clc1ccc(CNC(=N)NC(=N)NCCCCCCCC)cc1Cl.O=C(O)[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO

(±)-SIPI 6360

D2/5-HT2A receptor dual antagonist

7-[3-[4-(6-fluoro-1,2-benzoxazol-3-yl)piperidin-1-yl]propoxy]-3-methyl-3,4-dihydro-1H-quinolin-2-one

2(1H)-Quinolinone, 7-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy]-3,4-dihydro-3-methyl-

2(1H)-Quinolinone, 7-[3-[4-(6-fluoro-1,2-benzisoxazol-3-yl)-1-piperidinyl]propoxy]-3,4-dihydro-3-methyl-

((±)-SIPI 6360)

7-(3-(4-(6-Fluorobenzo[d]isoxazol-3-yl)piperidin-1-yl)propoxy)-3-methyl-3,4-dihydroquinolin-2(1H)-one

((±)-SIPI 6360)

Mp 154–155 °C;

¹H NMR (400 Hz, CDCl₃) δ 8.41 (br, 1H), 7.70 (dd, J = 8.8, 5.2 Hz, 1H), 7.27–7.23 (m, 1H), 7.08–7.03 (m, 2H), 6.53 (dd, J = 8.0, 2.4 Hz, 1H), 6.37 (d, J = 2.4 Hz, 1H), 4.02 (t, J = 6.0 Hz, 2H), 3.11–3.08 (m, 3H), 2.94–2.92 (m, 1H), 2.67–2.56 (m, 4H), 2.18–2.00 (m, 8H), 1.28 (d, J = 6.4 Hz, 3H);

¹³C NMR (100 MHz, CDCl₃) δ 174.61, 164.10 (d, J = 249.0 Hz), 163.86 (d, J = 14.0 Hz), 161.11, 158.59, 138.06, 128.74, 122.59 (d, J = 11.0 Hz), 117.30, 115.71, 112.31 (d, J = 25.0 Hz), 108.45, 101.96, 97.44 (d, J = 27.0 Hz), 66.43, 55.38, 53.62, 35.25, 34.61, 32.68, 30.55, 26.88, 15.34;

MS m/z 437.6 [M + H]⁺;

HRMS (ESI) m/zcalcd for C₂₅H₂₉FN₃O₃ [M + H]⁺ 438.2193, found 438.2210.

Synthesis

Schizophrenia is a common severe mental patients, mental illness is the most serious of all, the most dangerous kind, the worldwide incidence of about I%, with the accelerate pace of social life, The incidence was significantly increased. Most schizophrenic patients due to the long treatment period, high cost, side effects and give up the treatment, often lead to more serious social consequences.

Numerous studies show that the brain monoamine neurotransmitters, especially dopamine and 5-hydroxytryptamine system is closely related to the body’s normal mental activity, these two types of system disorder can lead to a variety of neuropsychiatric diseases such as schizophrenia , neuropathic pain, mania, anxiety disorders, all kinds of depression, Parkinson’s disease and the like.

The drugs currently used clinically primarily for conventional antipsychotics (such as dopamine D2 receptor antagonists) and atypical antipsychotics (such as D2 / 5-HT2a dual antagonist), where conventional antipsychotics because it is easy leads to extrapyramidal symptoms (EPS) and gradually phased out, atypical antipsychotics variety, but no one medication to improve the overall spectrum of schizophrenia has the absolute advantage, most of the positive or negative symptoms of a a symptom improvement, or reduced side effects. So look for low toxicity, rapid onset, treatment spectral width of new anti-schizophrenia drug has been a hot topic in the world pharmaceutical industry.

In recent years, scientists have found that the dopamine D2 partial agonist can over time reduce dopamine activity in the transfer of dopamine, but not all block; the other hand, when the low dopaminergic activity is caused by stimulating effect on both positive and negative symptoms of mental illness have a significant effect. 5-HT2a receptor antagonists can improve negative symptoms, while synergies D2 EPS side effects can be reduced to about 1% level (classical antipsychotic drugs EPS incidence is about 30%), part of the 5-HTla agonism and 5-HT2a and synergy can make in therapeutic doses under observation EPS decreased to undetectable levels, therefore, has D2 ,5-HT2a, 5HTla synergy targets three new anti-drugs are currently developed Jingshenfenlie focus and an important development direction.

The present invention relates to a quinoline derivative can stabilize the brain dopaminergic, serotonergic energy system, may for a variety of neurological and psychiatric diseases have improved and treatment can be used for neuropathic pain, mania, schizophrenia, anxiety disorders, all kinds of depression, Parkinson’s disease, especially in the treatment of schizophrenia.

DETAILS COMING……….

PATENT

CN 102718758

PATENT

WO 2012130153

Example 1

1-1

7- (3- (4- (6-fluorophenyl and [d] different dumb-3-yl) piperidin-1-yl) propoxy) -3,4-dihydro-3-carboxylic acid -one – yl quinolin -2 (1H)

1) N- (3- methoxyphenyl) propionamide

3-methoxy-aniline (0.1mol), methylene chloride (30 mL), triethylamine (0.2mol), was added to the flask lOOmL three, propionyl chloride was added dropwise under ice (0.12mol) in methylene chloride 30 mL, temperature does not exceed 5 ° C, the addition was complete, the ice bath was removed and stirred at room temperature 0.5h, the system was washed with water, dilute hydrochloric acid, saturated brine, dried over anhydrous magnesium sulfate, and evaporated to dryness to give a white powdery solid 17.01g yield 95%.

2) 2-chloro-7-methoxy-3-methylquinoline

The DMF (20mL) was added to the three 250mL flask, was added dropwise under ice-salt bath of POCl ₃ (100 mL), temperature does not exceed 0 ° C, the addition was completed stirring 0.5h, was added portionwise N- (3- methoxyphenoxy yl) propanamide powder (31.0g), was slowly warmed to 50 ° C, violent reaction, to be exothermic easing slowly warmed to reflux, the reaction was kept 2h, cooled to room temperature, the system was poured into 800 g of crushed ice to sodium carbonate to adjust the pH to 7 to precipitate a yellow solid with petroleum ether – ethyl acetate to give pure product 20.86g, yield 58%.

3) 3-methyl-7-methoxy-quinolin -2 (1H) – one

2-Chloro-7-methoxy-3-methyl-quinoline (20.76g), acetic acid (150 mL) placed in 250mL one-neck flask, heated at reflux for 24h, acetic acid recovery, and the residue was recrystallized from ethanol to 95%, white needle crystalline 16.08g, yield 85%.

4) 7-methoxy-3,4-dihydro-3-methyl-quinolin -2 (1H) – one

7-Methoxy-3-methyl-quinolin -2 (1H) – one (18.92g), acetic acid (150mL), 10% Pd / C (lg) was added to the three 250mL flask, the system was replaced with nitrogen air, and then the nitrogen was replaced with hydrogen, and then the reaction was heated to 80 ° C overnight, cooled to room temperature, filtered and the filtrate evaporated to dryness to give a white powder, washed with water once, 50 ° C and dried in vacuo 4h, as a white powdery solid 18.91g yield of 98.95%.

5) 7-hydroxy-3,4-dihydro-3-methyl-quinolin -2 (1H) – one

7-Methoxy-3,4-dihydro-3-methyl-quinolin -2 (1H) – one (19.12g), 40% hydrobromic acid (150 mL) placed in 250mL one-neck flask was heated at reflux for 12h cooled to room temperature, the precipitated solid was filtered, the filter cake successively with hydrobromic acid, washed with water, 50 ° C and dried in vacuo 4h, 14.60 g as a white powdery solid, yield 82.4%.

6) 3- (1- (3-chloropropyl) piperidin-4-yl) -6-fluorophenyl and [d] oxazole different dumb

6-fluoro-3- (piperidin-4-yl) benzo [d] isoxazol dummy oxazole (22.00g), 1- bromo-3-chloropropane (40mL), anhydrous potassium carbonate (40g), acetone ( 250mL) was added to a 500mL one-neck flask was refluxed overnight, cooled to room temperature, filtered, the filter cake was washed twice with hot acetone and the combined filtrate was added dropwise a solution of anhydrous hydrogen chloride in ethanol, the precipitated white solid was filtered, the filter cake washed with acetone after washing once, it was dissolved in 200mL of water, adjusted with sodium carbonate to pH 9, and filtered to obtain a white powdery solid 15.94 g, yield 48.0%

7) 7- (3- (4- (6-fluorobenzo [d] isoxazol-3-yl dummy) piperidin-1-yl) propoxy) -3,4-dihydro-3-methyl quinolin -2 (1H) – one

3- (1- (3-chloropropyl) piperidin-4-yl) -6-fluorophenyl and [d] oxazole different dumb (lmmol), 7- hydroxy-3,4-dihydro-3-carboxylic acid yl quinolin -2 (1H) – one (1.0 mmol), anhydrous potassium carbonate (3.0mmol) were added to the lOmLDMF, 60 ° C overnight the reaction, potassium carbonate was filtered off, the mother liquor evaporated to dryness to give a pale yellow solid, the filter cake recrystallized with 95% ethanol, 50 ° C and dried in vacuo 4h, as a white powdery solid 0.30g, 69% yield.

NMR IH (of DMSO-D ^{. 6} ): L up to .27 (D, 3H, J = 9.2Hz), 2.06-2.32 (m, 9H), 2.67-2.69 (T, 2H), 2.95 (D * D, lH, J = 3.2Hz, 12.8Hz), 3.15-3.17 ( m, 2H), 4.05 (t, 2H, J = 6Hz), 6. 37 (d, lH, J = 2.4Hz), 6.56 (d * d, lH, J = 2.4Hz, 8.0Hz), 7.05-7.11 (m, 2H), 7.25-7.29 (m, lH), 7.73-7.76 (m, lH), 7.98 (s, lH), 11.43 (brs, lH)

ESI-MS: 438 (M + 1)

Example 2

Preparation 1-1 hydrochloride

7- (3- (4- (6-fluorophenyl and [d] different dumb-3-yl) piperidin-1-yl) propoxy) -3,4-dihydro-3-methyl-quinoline morpholine -2 (1H) – one (lmmol) was dissolved with ethyl acetate (50 mL) was slowly added dropwise a solution of anhydrous hydrogen chloride in ethyl acetate (lmol / L, 5mL), stirred for 2h, the precipitated solid was filtered, the filter cake washed with ethyl acetate, 50 ° C and dried in vacuo 4h, as a white powdery solid 0.436g, yield 92%.

ESI-MS: 438 (M + 1)

Elemental analysis results:

Calcd: C, 63.35%; H, 6.17%; Cl, 7.48%; F, 4.01%; N, 8.87%; O, 10.13%

Found: C, 63.29%; H, 6.24%; CI, 7.43%; F, 4.05%; N, 8.82%; O, 10.17%

Example 3

Preparation 1-1 methanesulfonate

The 1-1 (lmmol) was dissolved with ethyl acetate (50 mL) was slowly added dropwise a solution of methanesulfonic acid in ethyl acetate (lmol / L, 5mL), stirred for 2h, the precipitated solid was filtered, the filter cake with ethyl acetate wash, 50 ° C and dried in vacuo 4h, as a white powdery solid 0.487g, yield 91.3%.

ESI-MS: 438 (M + 1, positive mode), 95 (CH ₃ the SO ₃ -, negative mode) Elemental analysis:

Calcd: C, 58.52%; H, 6.04%; F, 3.56%; N, 7.87%; 0, 17.99%; S, 6.01%

Found: C, 58.49%; H, 6.09%; F, 3.50%; N, 7.81%; 0, 18.02%; S, 6.09%

PATENT

US 20110160199

Paper

Development and Kilogram-Scale Synthesis of a D₂/5-HT_2A Receptor Dual Antagonist (±)-SIPI 6360

The kilogram-scale synthesis of a D₂/5-HT_2A receptor dual antagonist (±)-SIPI 6360 was developed as an alternative treatment for schizophrenia. Specifically, three conditions were modified and optimized, including the Vilsmeier conditions, to prepare quinoline 3. In addition, the palladium-catalyzed hydrogenation was modified to synthesize dihydroquinolin-2(1H)-one 5, and the reduction of β-chloroamide was altered to form 3-chloropropanamine 8. Ultimately these improvements led to the preparation of a 1.5 kg of (±)-SIPI 6360 batch in eight steps with an overall yield of 34% and purity of 99.8%.

//////// D2/5-HT2A receptor dual antagonist (±)-SIPI 6360, 1401333-14-9

c21CC(C(Nc1cc(cc2)OCCCN3CCC(CC3)c4c5ccc(cc5on4)F)=O)C