Saturday 9 May 2015

Daiichi Sankyo Co. v. Matrix Laboratories, Ltd. (Fed. Cir. 2010)

Daiichi Sankyo



In Daiichi Sankyo Co. v. Matrix Laboratories, Ltd., decided last Thursday, the Federal Circuit affirmed a determination by the District Court for the District of New Jersey that Matrix Laboratories, Ltd., Mylan Inc., Mylan Laboratories, Inc., and Mylan Pharmaceuticals, Inc. ("Mylan") failed to establish a prima facie case of obviousness with respect to claim 13 of U.S. Patent No.5,616,599.  Daiichi Sankyo Company, Ltd. and Daiichi Sankyo, Inc. own the '599 patent, which relates to 1-biphenylmethylimidazole compounds and their use as angiotensin receptor blockers (ARBs) for the treatment of high blood pressure.  Claim 13 of the '599 patent encompasses olmesartan medoxomil, an ARB that is marketed by Daiichi as the active ingredient in Benicar®, Benicar HCT®, and Azor®.
The use of ARBs to control blood pressure can be traced back to work done in the 1970s and 1980s by Takeda Pharmaceutical Co. Ltd., which developed a class of compounds comprising an imidazole ring with other chemical moieties bonded to the five positions of the ring.  One such compound, Takeda's S-8307, has the chemical structure:
S-8307
Using Takeda's compounds as leads, E. I. du Pont de Nemours and Company sought to develop compounds having inreased angiotensin receptor binding, and therefore, better ARB activity.  DuPont modified S-8307 to develop the compound losartan, which has ten-fold higher binding than Takeda's compounds.  DuPont's losartan has the chemical structure:
Losartan
DuPont disclosed losartan in U.S. Patent No. 5,138,069 along with more than 400 structurally related ARBs, including Example 118, which has the chemical structure:
Example 118
Based on DuPont's success with losartan, a number of pharmaceutical companies, including Daiichi, initiated efforts to identify even better ARBs.  Daiichi's work led to the synthesis of olmesartan, the active metabolite of olmesartan medoxomil, which differs from losartan in that it has a hydrophilic, hydroxy-isopropyl group at the 4-position of the imidazole ring instead of a lipophilic, chlorine atom (like Example 118 above), and a carboxy group masked by a medoxomil prodrug substituent at the 5-position of the ring instead of a hydroxymethyl group.  Olmesartan medoxomil and olmesartan have the chemical structures:



Olmesarton
























The closest prior art structure to Daiichi's olmesartan is DuPont's Example 6, disclosed in U.S. Patent No. 5,137,902, which differs from olmesartan in that it lacks a single oxygen atom at the 4-position of the imidazole ring.  DuPont's Example 6 has the chemical structure (wherein the circled hydrogen atom in Example 6 is an -OH in olemsartan):
Example 6
Matrix Laboratories





































Seeking approval to market generic olmesartan medoxomil, Mylan filed multiple ANDAs with the FDA.  In response to Mylan's ANDA filings, Daiichi brought suit against Mylan for infringement of claim 13 of the '599 patent.  The parties stipulated to infringement, leaving Mylan's counterclaim that claim 13 would have been obvious in light of:  (1) the ARBs disclosed in DuPont's '902 patent, (2) Example 118 in DuPont's '069 patent, and (3) the well-known use of medoxomil as a prodrug.  In particular, Mylan contended that one of skill in the art would have been motivated to select the ARBs disclosed in DuPont's '902 patent as lead compounds, and then modify the lipophilic alkyl groups at the 4-position of those compounds with olmesartan's hydrophilic hydroxyalkyl group in view of Example 118.
Following a bench trial, the District Court held that claim 13 of the '599 patent was not invalid as obvious, finding that Mylan had failed to show by clear and convincing evidence that a skilled artisan would have chosen the ARBs disclosed in DuPont's '902 patent as lead compounds, that the structure of the '902 patent compounds differed significantly from olmesartan medoxomil, and that Mylan had failed to prove that a skilled artisan would have been motivated to modify the 4- and 5-positions of the '902 patent compounds to obtain olmesartan medoxomil.  In particular, the District Court determined that the '069 patent and its ARBs taught away from the use of a hydrophilic group at the 4-position and from any expectation that the use of a hydrophilic group would generate an ARB with significantly improved biological properties, and further, that converting olmesartan into a prodrug was a disfavored and unpredictable approach and that medoxomil was a disfavored prodrug.  The District Court also found that secondary considerations favored a finding of nonobviousness; specifically that olmesartan medoxomil's enhanced potency constituted evidence of unexpected results and that Daiichi's Benicar® enjoyed commercial success despite being the seventh ARB on the market.


Mylan #1


Federal Circuit Seal




















































In affirming the District Court's finding of nonobviousness, the Federal Circuit agreed with Daiichi in determining that Mylan failed to show (1) that one of ordinary skill in the art would have been motivated to select the ARBs disclosed in DuPont's '902 patent as lead compounds or (2) that the skilled artisan would have been motivated to modify the '902 patent compounds to synthesize olmesartan medoxomil.  The Court began its analysis by citingEisai Co. Ltd. v. Dr. Reddy's Labs., Ltd., 533 F.3d 1353 (Fed. Cir. 2008), andTakeda Chem. Indus., Ltd. v. Alphapharm Pty., Ltd., 492 F.3d 1350 (Fed. Cir. 2007), for the proposition that:
Proof of obviousness based on structural similarity [between claimed and prior art compounds] requires clear and convincing evidence that a medicinal chemist of ordinary skill would have been motivated to select and then to modify a prior art compound (e.g., a lead compound) to arrive at a claimed compound with a reasonable expectation that the new compound would have similar or improved properties compared with the old.
With respect to the issue of lead compound selection, the panel countered Mylan's argument that because the '902 patent compounds are the closest prior art, this should have been dispositive of the lead compound issue, pointing out that such argument "runs contrary to our case law."  The Court noted that "[i]nTakeda, we upheld a district court's finding that one of skill in the art would not have chosen the structurally closest prior art compound, compound b, as the lead compound in light of other compounds with more favor-able characteristics," adding that the Court's cases "illustrate that it is the possession of promising useful properties in a lead compound that motivates a chemist to make structurally similar compounds."  Citing Ortho-McNeil Pharm., Inc. v. Mylan Labs., Inc., 520 F.3d 1358 (Fed. Cir. 2008), the Court explained that "attribution of a compound as a lead compound after the fact must avoid hindsight bias; it must look at the state of the art at the time the invention was made to find a motivation to select and then modify a lead compound to arrive at the claimed invention" (emphasis in original).  Thus, the selection of a lead compound "depends on more than just structural similarity, but also knowledge in the art of the functional properties and limitations of the prior art compounds," and therefore, "[p]otent and promising activity in the prior art trumps mere structural relationships."
Turning to the issue of motivation to modify, the panel noted that the vast majority of compounds disclosed in the '069 patent contain a lipophilic group at the 4-position of the imidazole ring, and that only four (Examples 342, 329, 118, and 335) have a hydrophilic group like olmesartan medoxomil.  According to the Court, "[t]he few compounds with hydrophilic groups at the 4-position are drowned out by the sea of 4-lipophilic compounds, which are the essence of what the ’069 patent teaches."  In addition, the Court observed that binding affinity analyses comparing the '069 patent compounds that differed only at the 4-position confirm the preference for lipophilicity at that position.  The panel determined that:
Altogether, the '069 patent's [structural-activity relationship] data and the structure of other second-generation ARBs counter any notion that one of skill in the art would have been motivated to modify the '902 compounds' lipophilic alkyl groups to a hydrophilic group.  Such a holding would have been based on hindsight.
The panel therefore affirmed both the District Court's finding that Mylan had failed to establish that a skilled artisan would have selected the '902 patent ARBs as lead compounds and the lower court's finding that a skilled artisan would have modified the '902 patent ARBs at the 4-position of the imidazole ring to obtain olmesartan medoxomil.  As a result, the Federal Circuit affirmed the District Court's determination that claim 13 of the '599 patent was not shown to be invalid as obvious.
Daiichi Sankyo Co. v. Matrix Laboratories, Ltd. (Fed. Cir. 2010)
Panel: Circuit Judges Lourie, Friedman, and Linn
Opinion by Circuit Judge Lourie

BIOCON PATENT..........US20140288269) PROCESS FOR THE PREPARATION OF RANDOM POLYPEPTIDES AND EMPLOYING CIRCULAR DICHROISM AS A GUIDANCE TOOL FOR THE MANUFACTURE OF GLATIRAMER ACETATE .




PROCESS FOR THE PREPARATION OF RANDOM POLYPEPTIDES AND EMPLOYING CIRCULAR DICHROISM AS A GUIDANCE TOOL FOR THE MANUFACTURE OF GLATIRAMER ACETATE (Fri, 26 Sep 2014) 
CLICK read this at wipo


OWNER

BANGALORE: Biotech major Biocon today said its Chairperson and Managing Director Kiran Mazumdar-Shaw











Application Number:14355504Application Date:26.10.2012
Publication Number:20140288269Publication Date:25.09.2014
Publication Kind :A1
PCT Reference: Application Number:PCT/IN2012/000708 ; Publication Number: Click to see the data
IPC:
C07K 1/04
C07K 1/14
Applicants:Biocon Limited
Inventors:Venkata Srinivas Pullela
Khedkar Anand
Patil Nitin Sopanrao
Ujire Sandhya
Chatterjee Amarnath
Janakiraman Ashwini
Priority Data:3781/CHE/2011 03.11.2011 IN
Title:(EN) PROCESS FOR THE PREPARATION OF RANDOM POLYPEPTIDES AND EMPLOYING CIRCULAR DICHROISM AS A GUIDANCE TOOL FOR THE MANUFACTURE OF GLATIRAMER ACETATE
Abstract:
(EN)
The present invention discloses novel process for the preparation of mixture of polypeptides comprising L-Glutamaic acid, L-Alanine, L-Tyrosine, and L-Lysine. by employing circular dichroism as a guidance tool.


Glatiramer is a peptide based polymer composed of four amino acids: L-Glutamaic acid, L-Alanine, L-Tyrosine, and L-Lysine. It's pharmaceutically acceptable salt Glatiramer acetate is approved by FDA and marketed as Copaxone® for the treatment of multiple sclerosis. Copaxone is also known as copolymer-1 and cop-1. Multiple sclerosis is an autoimmune disease affects the brain and central nervous system due to the damage to the myelin sheath of the nerve cells, which results as demyelination of axons. Glatiramer acetate is a synthetic polypeptide analogue of myelin basic protein (MBP). Pharmacologically, Copaxone is a non-interferon and non-steroidal immunomodulator, which arrests the multiple sclerosis aggression. Glatiramer acetate is administrated by subcutaneous injections.
      Chemically, glatiramer acetate is designated L-glutamic acid polymer with L-alanine, L-lysine and L-tyrosine, acetate salt. Its structural formula is:

          (Glu, Ala, Lys, Tyr)x.xCH3COOH 
          (C5H9NO4.C3H7NO2.C6H14N2O2.C9H11NO3)x.xC2H4O2
      Average molecular weight of glatiramer acetate is 5,000-11,000 daltons and the average molar fractions of the respective amino acids are: 0.141, 0.427, 0.095, and 0.338.
      U.S. Pat. No. 3,849,550 describes the observation of MBP arrest in experimental allergic encephalomyelitis (a disease similar to multiple sclerosis) by immunotherapy agents. With continuous endeavours, glatiramer acetate is resulted as an advanced analogue for the treatment of multiple sclerosis with improved safety and efficacy.
      U.S. Pat. Nos. 5,800,808; 5,981,589; 6,048,898 describes the process preparation of glatiramer acetate employing the N-carboxyanhydrides (NCAs) derived from alanine, γ-benzyl glutamate, N-trifluoroacetyl lysine, and tyrosine. Following the steps: polymerization, sequential cleavage of the γ-benzyl ester of glutamate and N-trifluoroacetyl derivative of lysine, acetate salt formation and final purification. U.S. Pat. No. 6,620,847 describes a process for the preparation of glatiramer acetate using the aqueous piperidine for trifluoroacetyl cleavage of lysine. U.S. Pat. No. 7,049,399 describes the process for preparation of polypeptide-1 using the catalytic transfer hydrogenation for the cleavage of γ-benzyl ester of glutamate. E.P. Pat. No. 1,807,467 describes the processes for preparation of glatiramer using NCAs of alanine, tyrosine, N-t-butoxycarbonyl L-Lysine, and protected glutamic acid, where in the protecting group is selected from γ-methoxybenzyl and γ-benzyl. U.S. Pat. No. 7,495,072 describes the process for the preparation of mixtures of polypeptides using purified hydrobromic acid. The major drawback of all these processes is the generation of impurities, multiple steps of purification and the variability in the secondary structures amongst different batches manufactured using the same process.

HO

Map of Biocon



  1. Biocon
  2. Pharmaceutical Company
  3. Address: Door No. A - 210, Gokul Arcade, Garevan Chowk, Sahar Road, Vile Parle East, Mumbai - 400057
    Phone:022 6691 9761


EXAMPLES

Example 1

Preparation of Glatiramer Acetate Using Potassium Tert Butoxide

Preparation of Protected Polymer 2

      Protected copolymer (1 gm) was taken in a mixture of THF and water, to that Lewatit K 2629 resin (1 gm) was added and stirred at 65° C. for 24 h. The resin was filtered through buckner funnel and washed with THF (5 ml).The reaction mass was distilled to 3-4 volume stage and water was added and the precipitated product was filtered and dried in VTD for 24 h at 40-45° C. Yield: 0.6 gm

Preparation of Glatiramer Acetate

      To the stirred solution of protected polymer 2 (0.6 g) in anhydrous methanol (9 ml) was added potassium tertiary butoxide (0.6 g) and stirred for 1 hour. Reaction mass was concentrated under reduced pressure (below 35 C.). To the reaction mass water (0.6 mL) was added and pH was adjusted with Glacial acetic acid to 5.5. Crude glatiramer acetate was isolated by crystallising with acetone. Crystallised solid was filtered and suck dried. Yield: 0.4 g
      Crude glatiramer acetate obtained is subjected for gel permeation chromatography for purification.

Example 2

Preparation of Glatiramer Acetate Using Sodium Methoxide

Preparation of Protected Polymer 2

      Protected copolymer (1 gm) was taken in a mixture of THF (8 ml) and water (ml), to that Lewatit K 2629 resin (1 gm) was added and stirred at 65° C. for 24 h. The resin was filtered through buckner funnel and washed with THF (5 ml).The reaction mass was distilled to 3-4 volume stage and water was added and the precipitated product was filtered and dried in VTD for 24 h at 40-45° C. Yield: 0.6 gm

Preparation of Glatiramer Acetate

      To the stirred solution of protected polymer 2 (0.6 g) in anhydrous methanol (6 ml) was added a solution of sodium methoxide (0.9 g) in anhydrous methanol (4.5 mL)and stirred for 7 hours. Reaction mass was concentrated under reduced pressure (below 35° C.). To the reaction mass water (0.6 mL) was added and pH was adjusted with Glacial acetic acid to 6. Crude glatiramer acetate was isolated by crystallising with acetone. Crystallised solid was filtered and suck dried. Yield: 0.4 g
      Crude glatiramer acetate obtained is subjected for gel permeation chromatography for purification.

Example 3

Preparation of Glatiramer Acetate Using TMSCl/NaI Followed by Sodium Methoxide

      Protected polymer 1 (20 g) was charged in THF (200 ml) under nitrogen atmosphere, added sodium iodide (1 g) was added followed by trimethylsilyl chloride (20 ml) at room temperature and stirred for 3 h. The reaction mass was quenched after the completion of reaction with water (20 ml). The solids were filtered, washed with water (100 ml) and dried under high vacuum to obtain protected copolymer 2 (10 g).
      The resulted protected polymer 2 was suspended in anhydrous methanol (100 ml), solution of sodium methoxide (15 g)in anhydrous methanol (75 ml) was added and stirred at room temperature for 7 h. pH was adjusted after the completion of the reaction to 6 with glacial acetic acid, and the mass was purified to obtain glatiramer acetate (6 g).

Example 4

Preparation of Glatiramer Acetate from Protected Polymer 3

      Solution of sodium methoxide (1.5 g) in anhydrous methanol (7.5 ml) was added to protected copolymer 3 (1 g) in anhydrous methanol (10 ml) at room temperature and stirred for 7 h. pH was adjusted to 5 after completion of the reaction with glacial acetic acid. The resulted mass was purified to obtain glatiramer acetate (0.6 g).
      The Far UV CD spectra, of Glatiramer acetate synthesised using the processes described in examples 1-4 exhibits the presence of random coils in the wavelength region 195-215 nm and alpha helices in the wavelength region of 222 nm. (FIG. 5)










OWNER

BANGALORE: Biotech major Biocon today said its Chairperson and Managing Director Kiran Mazumdar-Shaw




BANGALORE

BENGALURU, INDIA,


Map of bangalore



Image result for BANGALORE


Image result for bangalore metro


























BULL TEMPLE










Thursday 7 May 2015

WO 2015059679 New patent on Eliglustat by Dr Reddys

WO-2015059679

 
Eliglustat tartrate (Genz-1 12638) is a glucocerebroside (glucosylceramide) synthase inhibitor for the treatment of gaucher disease and other lysosomal storage disorders, which is currently under development.
Eliglustat is chemically known as 1 R, 2R-Octanoic acid [2-(2', 3'-dihydro-benzo [1 , 4] dioxin-6'-yl)-2-hydroxy-1 -pyrrolidin-1 -ylmethyl]-ethyl]-amide, having a structural formula I depicted here under.
Formula I
Eliglustat hemitartrate (Genz-1 12638) development by Genzyme, is a glucocerebroside (glucosylceramide) synthase inhibitor for the treatment of Gaucher disease and other lysosomal storage disorders. Eliglustat hemitartrate is orally active with potent effects on the primary identified molecular target for type 1 Gaucher disease and other glycosphingolipidoses, appears likely to fulfill high expectations for clinical efficacy. 

Gaucher disease belongs to the class of lysosomal diseases known as glycosphingolipidoses, which result directly or indirectly from the accumulation of glycosphingolipids, many hundreds of which are derived from glucocerebroside. The first step in glycosphingolipid biosynthesis is the formation of glucocerebroside, the primary storage molecule in Gaucher disease, via glucocerebroside synthase (uridine diphosphate [UDP] - glucosylceramide glucosyl transferase). Eliglustat hemitartrate is based on improved inhibitors of glucocerebroside synthase.

U.S. patent No. 7,196,205 (herein described as US'205) discloses a process for the preparation of eliglustat or a pharmaceutically acceptable salt thereof. In this patent, eliglustat was synthesized via a seven-step process involving steps in that sequence: 
(i) coupling S-(+)-2-phenyl glycinol with phenyl bromoacetate followed by column chromatography for purification of the resulting intermediate, 
(ii) reacting the resulting (5S)-5-phenylmorpholin-2-one with 1 , 4-benzodioxan-6-carboxaldehyde to obtain a lactone, 
(iii) opening the lactone of the oxazolo-oxazinone cyclo adduct via reaction with pyrrolidine, 
(iv) hydrolyzing the oxazolidine ring, (v) reducing the amide to amine to obtain sphingosine like compound, (vi) reacting the resulting amine with octanoic acid and N-hydroxysuccinimide to obtain crude eliglustat, (vii) purifying the crude eliglustat by repeated isolation for four times from a mixture of ethyl acetate and n-heptane.

U.S. patent No. 6855830, 7265228, 7615573, 7763738, 8138353, U.S. patent application publication No. 2012/296088 disclose processes for preparation of eliglustat and intermediates thereof.

U.S. patent application publication No. 2013/137743 discloses (i) a hemitartrate salt of eliglustat, (ii) a hemitartrate salt of eliglustat, wherein at least 70% by weight of the salt is crystalline, (iii) a hemitartrate salt of Eliglustat, wherein at least 99% by weight of the salt is in a single crystalline form.

https://patentscope.wipo.int/search/en/detail.jsf;jsessionid=234E6BE008E68831F6875FB703760826.wapp2nA?docId=WO2015059679&recNum=1&office=&queryString=FP%3A%28dr.+reddy%27s%29&prevFilter=%26fq%3DCTR%3AWO&sortOption=Pub+Date+Desc&maxRec=364


http://newdrugapprovals.org/2015/05/08/eliglustat/



WO-2015059679
Enhanced
Process for the preparation of eliglustat free base - comprising the reaction of S-(+)-phenyl glycinol with phenyl-alpha-bromoacetate to obtain 5-phenylmorpholin-2-one, which is further converted to eliglustat.



Dr Reddy's Laboratories Ltd




New crystalline eliglustat free base Form R1 and a process for its preparation are claimed. Also claimed is a process for the preparation of eliglustat free base which comprises the reaction of S-(+)-phenyl glycinol with phenyl-alpha-bromoacetate to obtain 5-phenylmorpholin-2-one, which is further converted to eliglustat. 

Further eliglustat oxalate, its crystalline form, and a process for the preparation of crystalline eliglustat oxalate, are claimed.

Formula II






http://newdrugapprovals.org/2015/05/08/eliglustat/



Example 1 : Preparation of 5-phenyl morpholine-2-one hydrochloride
To a (S) + phenyl glycinol (100g) add N, N-diisopropylethylamine (314ml) and acetonitrile (2000ml) under nitrogen atmosphere at room temperature. It was cooled to 10- 15° C. Phenyl bromoacetate (172.4g) dissolved in acetonitrile (500ml) was added to the above solution at 15° C over a period of 30 min. The reaction mixture is allowed to room temperature and stirred for 16-20h. Progress of the reaction was monitored by thin layer chromatography. After completion of the reaction, the reaction mixture was concentrated under reduced pressure at a water bath
temperature less than 25° C to get a residue. The residue was dissolved in ethyl acetate (1000ml) and stirred for 1 h at 15-20°C to obtain a white solid. The solid material obtained was filtered and washed with ethyl acetate (200ml). The filtrate was dried over anhydrous sodium sulphate (20g) and concentrated under reduced pressure at a water bath temperature less than 25° C to give crude compound (1000g) as brown syrup. The Crude brown syrup is converted to HCI salt by using HCI in ethyl acetate to afford 5-phenyl morpholine-2-one hydrochloride (44g) as a white solid. Yield: 50%, Mass: m/z = 177.6; HPLC (% Area Method): 90.5%


Example 2: Preparation of (1 R,3S,5S,8aS)-1 ,3-Bis-(2',3'-dihydro-benzo[1 ,4] dioxin-6'-yl)-5-phenyl-tetrahydro-oxazolo[4,3-c][1 ,4]oxazin-8-one.
5-phenyl morpholine-2-one hydrochloride (100g) obtained from above stage 1 is dissolved in toluene (2500ml) under nitrogen atmosphere at 25-30°C. 1 ,4-benzodioxane-6-carboxaldehyde (185.3g) and sodium sulphate (400g) was added to the above solution and the reaction mixture was heated at 100-105°C for 72h. Progress of the reaction was monitored by thin layer chromatography. After completion of reaction, the reaction mixture was concentrated under reduced pressure at a water bath temperature less than 25° C to get a residue. The residue was cooled to 10°C, ethyl acetate (2700ml) and 50% sodium bisulphate solution (1351 ml) was added to the residue and stirred for 1 h at 10°C to obtain a white solid. The obtained white solid was filtered and washed with ethyl acetate. The separated ethyl acetate layer was washed with water (1000ml), brine (1000ml) and dried over anhydrous sodium sulphate. The organic layer was concentrated under reduced pressure at a water bath temperature of 45-50°C to get a crude material. The obtained crude material is triturated with diethyl ether (1500ml) to get a solid material which is filtered and dried under vacuum at room temperature for 2-3h to afford (1 R,3S,5S,8aS)-1 ,3-Bis-(2',3'-dihydro-benzo[1 ,4]dioxin-6'-yl)-5-phenyl-tetrahydro-oxazolo[4,3-c][1 ,4]oxazin-8-one (148g) as a yellow solid. Yield: 54%, Mass: m/z = 487.7; HPLC (% Area Method): 95.4 %

Example 3: Preparation of (2S,3R,1 "S)-3-(2',3'-(Dihydro-benzo[1 ,4]dioxin-6'-yl)-3-hydroxy-2-(2"-hydroxy-1 ''^henyl-ethy^
(1 R,3S,5S,8aS)-1 !3-Bis-(2'!3'-dihydro-benzo[1 ,4]dioxin-6'-yl)-5-phenyl-tetrahydro-oxazolo[4,3-c][1 ,4]oxazin-8-one (70g) obtained from above stage 2 was dissolved in chloroform (1400ml) at room temperature. It was cooled to 0-5°C and pyrrolidone (59.5ml) was added at 0-5°C over a period of 30 minutes. The reaction mixture was allowed to room temperature and stirred for 16-18h. Progress of the reaction was monitored by thin layer chromatography. After completion of reaction, the reaction mixture was concentrated under reduced pressure at a water bath temperature of 40-45°C to obtain a crude. The obtained crude was dissolved in methanol (1190ml) and 1 N HCI (1 190ml) at 10-15° C, stirred for 10 minutes and heated at 80-85°C for 7h. Progress of the reaction was monitored by thin layer chromatography. After completion of reaction, methanol was concentrated under reduced pressure at a water bath temperature of 50-55°C.The aqueous layer was extracted with ethyl acetate and the organic layer was washed with 1 N HCI (50ml). The aqueous layer was basified with saturated sodium bicarbonate solution up to pH 8-9 and extracted with ethyl acetate (3x70ml). The combined organic layers was washed with brine (100ml), dried over anhydrous sodium sulphate and concentrated under reduced pressure at a water bath temperature of 50-55°C to afford (2S,3R,1"S)-3-(2',3'-(Dihydro-benzo[1 ,4]dioxin-6'-yl)-3-hydroxy-2-(2"-hydroxy-1 "-phenyl-ethylamino)-1 -pyrrolidin-1 -yl-propan-1 -one (53g) as a yellow foamy solid. Yield: 90%, Mass: m/z = 412.7, HPLC (% Area Method): 85.1 %

Example 4: Preparation of (1 R,2R,1 "S)-1-(2',3'-(Dihydro-benzo[1 ,4]dioxin-6'-yl)2-hydroxy-2-(2"-hydroxy-1 '-phenyl-ethylamino)-3-pyrrolidin-1-yl-propan-1-ol.
(2S,3R,1 "S)-3-(2',3'-(Dihydro-benzo[1 ,4]dioxin-6'-yl)-3-hydroxy-2-(2"-hydroxy-1 "-phenyl-ethylamino)-1 -pyrrolidin-1 -yl-propan-1 -one (2.5g) obtained from above stage 3 dissolved in Tetrahydrofuran (106ml) was added to a solution of Lithium aluminium hydride (12.2g) in tetrahydrofuran (795ml) at 0°C and the reaction mixture was heated at 60-65°C for 10h. Progress of the reaction was monitored by thin layer chromatography. After completion of reaction, the reaction mixture was cooled to 5- 10°C and quenched in saturated sodium sulphate solution (100ml) at 5-10°C. Ethyl acetate was added to the reaction mass and stirred for 30-45 min. The obtained solid is filtered through celite bed and washed with ethyl acetate. Filtrate was dried over anhydrous sodium sulphate and concentrated under reduced pressure at a water bath temperature of 50°C to afford (1 R,2R, 1"S)-1 -(2',3'-(Dihydro-benzo[1 ,4]dioxin-6'-yl)2-hydroxy-2-(2"-hydroxy-1 '-phenyl-ethylamino)-3-pyrrolidin-1 -yl-propan-1 -ol (43.51 g) as a yellow gummy liquid. The crude is used for the next step without further purification. Yield: 85%, Mass: m/z = 398.7, HPLC (% Area Method): 77 %


Example 5: Preparation of (1 R, 2R)-2-Amino-1-(2', 3'-dihydro-benzo [1 , 4] dioxin-6'-yl)-3-pyrrolidin-1 -yl-propan-1 -ol.
(1 R,2R,1 "S)-1 -(2',3'-(Dihydro-benzo[1 ,4]dioxin-6'-yl)2-hydroxy-2-(2"-hydroxy-1 '-phenyl-ethylamino)-3-pyrrolidin-1 -yl-propan-1 -ol (40g) obtained from above stage 4 was dissolved in methanol (400ml) at room temperature in a 2L hydrogenation flask. Trifluoroacetic acid (15.5ml) and 20% Pd (OH) 2 (40g) was added to the above solution under nitrogen atmosphere. The reaction mixture was hydrogenated under H2, 10Opsi for 16-18h at room temperature. Progress of the reaction was monitored by thin layer chromatography. After completion of reaction, the reaction mixture was filtered through celite bed and washed with methanol (44ml) and water (44ml). Methanol was concentrated under reduced pressure at a water bath temperature of 50-55°C and the aqueous layer was washed with ethyl acetate. The aqueous layer was basified with 10M NaOH till the PH reaches 12-14 and then extracted with dichloromethane (2x125ml). The organic layer was dried over anhydrous sodium sulphate (3gm) and concentrated under reduced pressure at a water bath temperature of 45°C to obtain a gummy liquid. The gummy liquid was triturated with methyl tertiary butyl ether for 1 h to get a white solid, which is filtered and dried under vacuum at room temperature to afford (1 R, 2R)-2-Amino-1 -(2', 3'-dihydro-benzo [1 , 4] dioxin-6'-yl)-3-pyrrolidin-1 -yl-propan-1 -ol (23g) as a white solid. Yield: 82.3%, Mass (m/zj: 278.8, HPLC (% Area Method): 99.5%, Chiral HPLC (% Area Method): 97.9%


Example 6: Preparation of Eliglustat {(1 R, 2R)-Octanoic acid[2-(2',3'-dihydro-benzo [1 , 4] dioxin-6'-yl)-2-hydroxy-1 -pyrrolidin-1-ylmethyl-ethyl]-amide}.
(1 R, 2R)-2-Amino-1 -(2', 3'-dihydro-benzo [1 , 4] dioxin-6'-yl)-3-pyrrolidin-1 -yl-propan-1 -ol (15g) obtained from above stage 5 was dissolved in dry dichloromethane (150ml) at room temperature under nitrogen atmosphere and cooled to 10-15° C. Octanoic acid N-hydroxy succinimide ester (13.0 g)was added to the above reaction mass at 10-15° C and stirred for 15 min. The reaction mixture was stirred at room temperature for 16h-18h. Progress of the reaction was monitored by thin layer chromatography. After completion of reaction, the reaction mixture was cooled to 15°C and diluted with 2M NaOH solution (100 ml_) and stirred for 20 min at 20 °C. The organic layer was separated and washed with 2M sodium hydroxide (3x90ml).The organic layer was dried over anhydrous sodium sulphate (30g) and concentrated under reduced pressure at a water bath temperature of 45°C to give the crude compound (20g).The crude is again dissolved in methyl tertiary butyl ether (25 ml_) and precipitated with Hexane (60ml). It is stirred for 10 min, filtered and dried under vacuum to afford Eliglustat as a white solid (16g). Yield: 74%, Mass (m/zj: 404.7 HPLC (% Area Method): 97.5 %, ELSD (% Area Method): 99.78%, Chiral HPLC (% Area Method): 99.78 %.


Example 7: Preparation of Eliglustat oxalate.
Eliglustat (5g) obtained from above stage 6 is dissolved in Ethyl acetate (5ml) at room temperature under nitrogen atmosphere. Oxalic acid (2.22g) dissolved in ethyl acetate (5ml) was added to the above solution at room temperature and stirred for 14h. White solid observed in the reaction mixture was filtered and dried under vacuum at room temperature for 1 h to afford Eliglustat oxalate as a white solid (4g). Yield: 65.46%, Mass (m/zj: 404.8 [M+H] +> HPLC (% Area Method): 95.52 %, Chiral HPLC (% Area Method): 99.86 %



http://newdrugapprovals.org/2015/05/08/eliglustat/