ERlAL NUMBER 07/660,976 02
THIS APPLN IS A CON OF 07/384,187 07/21/39 ABN
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FILED SEPARATELY 07/384/187 07/21/89 548
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Foreign priority claimed 35 USC 119 conditionsmet yes Xno AS
JOAN THIERSTEIN a PATENT DEPARTMENT a WARNER-LAMBERT COMPANY 2 PLYMOUTH ROAD ANN ARBOR, MI 48105
LACTONE FORM AND SALTS THEREOF US. DEPT. of C0MM.-Pat. & TM Offlca - PTO-436L (rev. 10-78)
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INDEX OF CLAIMS
SYMBOLS Rejected Allowed (Through numeral) Canceled + Restricted N Non-elected I interference A Appeal Objected US005273995A United States Patent [19] [11] Patent Number: 5,273,995 . Roth [45] Date of Patent: Dec. 28, 1993
[54] [R-(R*R*)]-2-(4-FLOUROPHENYL)-B,-DIHY- [56] References Cited DROXY-5-(1-METHYLETHYL-3-PHENYl-4- U.S. PATENT DOCUMENTS (PHENYLAMINO) CARBONYL]- IH-PYRROLE-1-HEPTANOIC ACID, ITS 4,681,893 7/1987 Roth ...... 514/223 LACTONE FORM AND SALTS THEREOF OTHER PUBLICATIONS [75] Inventor: Bruce D. Roth, Ann Arbor, Mich. J. Med. Chem. 1985,28,347-358-G. E. Stokker, et al. “3-Hydroxy-3-Methylglutaryl-Coenzyme a Reduc- tase ... ”. [73] Assignee: Warner-Lambert Company, Morris Tetrahedron Letters, vol. 28, No. 13, pp. 1385-1388, Plains, N.J. 1987 “Synthesis of an HMG-COA Reductase Inhibitor, [21] Appl. No.: 660,976 Primary Eraminer-Mary C. Lee Assistant Examiner-Jacqueline Haley [22] Filed: Feb. 26, 1991 Attorney, Agent, or Finn-Ronald A, Daignault [57] ABSTRACT Related Application Data US. [R-(R*,R*)]-2-(4-fluorophenyl)-B,dihydroxy-5-((1- [63] Continuation of Ser. No. 384,187, Jul. 21, 1989, aban- methylethyl)-3-phenyl-4-[p@hcnylamino)carbonyl]- 1H- doned. pyrrole-1-heptanoic acid or (2R-trans)-5-(4-fluoro- phenyl)-2-(l-mcthylcthyl-N,4-diphcnyl-1-[2-(tctrahy- [51] Int. Cl.5 ...... A61K 31/40; C07D 405/06 dro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]- 1 H-pyrrole- [52] U.S. CI...... 514/422; 514/423; 3-carboxamide; and pharmaceutically acceptable salts 548/517; 548/537 thereof. [58] Field of Search ...... 514/422, 423; 548/517, 548/537 12 Claims, NO Drawings 5,273,995 1 2
[R*R*R*)]-2-(4-FLUOROPHENYL)-ß,§-DIHY- Ib DORXY-5-(1-METHYLETHYL-3-PHENYL-4- (PHENYLAMINO) CARBONYL]- 5 1H-PYRROLE-1-HEPTANOIC ACID, ITS LACTONE FORM AND SALTS THEREOF
07/384,187This is a filedcontinuation Jul. 21, 1989,of U.S. abandoned. application Ser. No. 10
BACKGROUND OF THE INVENTION Trans-(±)-5-(4-fluorophenyl)-2-(1-methylethyl)-N,4- diphenyl- 1-[2-tetrahydro-4-hydroxy-6-oxo-2H-pyran-2- 15 H yl)ethyl]-1H-pyrrole-3-carboxamides are among apparently is required for inhibition of HMG-GoA re- pounds of U.S. Pat. No. 4,681,893 having usefulness ps ductase. This is reported by Lynch et al. in “Synthesis inhibitors of cholesterol biosynthesis. The compounds of an HMB-CoA Reductase Inhibitor; A diastereoselec- therein broadly include 4-hydroxypayran-2-ones the 20 tive Aldol Approach in Tetrahedron Letters, Vol. 28, No. 13, pp. 1385-1388 (1987) as the 4R, 6R configura- corresponding ring-opened acids derived therefrom. tion. It is now unexpectedly found that the enantiomer However, an ordinarily skilled artisan may not pre- having the R form of the ring-opened acid of tran-5-(4- dict the unexpected and surprising inhibition of choles- fluorophcny1)-2-(11-mthylethyl-N-4-diphenyl-1 -1-[2-tet- 25 terol biosynthesis Of present invention in view Of these disclosures. rahydro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1 H- pyrrole-3-carboxamide; that is [R-(R*,R*)]-2-(4-fluoro- SUMMARY OF THE INVENTION phenyl)-ß,§-dihydroxy-5-(1 -methylethyl)-3-phenyl-4- Accordingly the present invention provides for com- [(phenylamino)carbonyl]-1H-pyrrole- l-heptanoic acid, 30 pounds consisting of [R-R*,R*)]-2-(4-fluorophenyl)- ß,§-dihydroxy-5-(( 1methylethyl)-3-phenyl-4- provides inhibition Of biosynthesis Of [(phenylamino) carbonyl]-1H-pyrrole-1-heptanoic acid cholesterol. (compound of formula I), pharmaceutically acceptable It is known that 3-hydroxy-3-methyIglutaryl coen- salts thereof and (2R-trans)-5-(4-fluorophenyl)-2-(1- zyme A (HMG-CoA) exists as the 3R-sterioisomer. 35 methylethyl-N,4-diphenyl-1-[2-tetrhydro-4-hydroxy- 1H-pyrrole-3-carboxamide Additionally, as shown in the study of a series of 5-sub- 6-oxo-2H-pyran-2-yl)ethyl] (the lactone of the heptanoic acid or of stituted 3,5-dihydroxypentanoic acids by Stokker et al., formula II). in “3-Hydroxy-3-methylglutaryl-Coenzyme A Reduc- The present invention also relates to a pharmaceutical tase Inhibitors. 1. Structural Modification of 5-Sub- 40 composition, useful a hypocholesterolemic agent, stituted 3,5-Dihydroxypentanoic acids and Their Lac- comprising a hypocholesterolemic effective amount of [R-(R*,R*)]-2-(4-fluorophenyl)-ß,§-dihydroxy-5(1- tone Derivatives,” J. Med. Chem. 1985, 28, 347-358, methylethyl)-3-phenyl-4-[(phenylamino)carbonyl]-H- essentially all of the biological activity resided in the pyrrole-1-heptanoic acid, its pharmaeutically accept- trans diastereomer of (E)-6-[2-(2,4-dichlorophenyl)e- 45 able salts Of (2R-trans)-5-(4-fluorohenyl)-2-(1- thenyl’-3,4,5,6-tetrahydro-4-hydroxy-2H-pyranone methylethyl-N,4-diphenyl- 1-[2-(tetrahydro-4-hydroxy- 6-oxo-2H-pyran-2-yl)thyl] 1H-pyrrole-3-carboxamide having a positive rotation. Further, the absolute config- acid; and a pharmaceutically acceptable Fur- uration for the ß-hydroxy-§-lactone moiety common to ther, the present invention is also a method of treating mevinolin of the formula (la) 50 mammals, including humans, suffering from hypercho- lesterolemia by administering to such mammal a dosage form of the pharmaceutical composition described above.
55 DETAILED DESCRIPTION OF THE INVENTION The pharmaceutically acceptable salts of the inven- tion are those generally derived by dissolving the free acid or the lactone; preferably the lactone, in aqueous or 60 aqueous alcohol solvent or other suitable solvents with an appropriate base and isolating the salt by evaporating the solution or by reacting the free acid or lactone; preferably the lactone and base in an organic solvent in which the salt separates directly or can be obtained by 65 concentration of the solution. H3C In practice, use of the salt form amounts to use of the acid or lactone form. Appropriate pharmaceutically and compactin of the formula (lb) acceptable salts within the scope of the invention are 5,273,995 3 4 those derived from bases such as sodium hydroxide, through a cationic exchange resin (+resin) and evap- potassium hydroxide, lithium hydroxide, calcium hy- orating the water. droxide, 11-deoxy-2-(methylamino)-D-glucitol, magne- The most preferred embodiment of the present inven- sium hydroxide, zinc hydroxide, aluminum hydroxide, tion is [R-(R*R*)]-2-(4-fluorophenyl-ß,§-dihydorxy-5- ferrous or ferric hydroxide, ammonium hydroxide or 5 (l-methylethyl)-3-phenyl-4((phenylamino)carbonyl]- organic amines such as N-methylglucamine, choline, IH-pyrrole- I-heptanoic acid, hemicalcium salt. arginine and the like. Preferably, the lithium, calcium, Generally, the compounds I and II of the present magnesium, aluminum and ferrous or ferric salts are invention can be prepared by (1) resolving the race- prepared from the sodium or potassium salt by adding mate, that is prepared by the process described in U.S. the appropriate reagent to a solution of the sodium or 10 Pat. No. 4,681,893 which is incorporated by reference potassium salt, i.e., addition of calcium chloride to a therefor, or (2) synthesizing the desired chiral form solution of the sodium or potassium salt of the com- beginning from starting materials which arc known or pound of the formula I will give the calcium salt readily prepared using processes analogous to those thereof. which are known. The free acid can be prepared by hydrolysis of the 15 Specifically, resolution of the racemate may be ac- lactone form of formula II or by passing the salt complished as shown in Scheme I (where Ph is phenyl) . as follows:
Scheme 1
PhHN PhHN
0 0 Ph Step A trans racemate +
R
OH OH 0
PhHN
0
1) Separate 2) NaOH 3) reflux in toluene
Ph CONHPh Ph CONHPh [R(R*R*) isomer [S(R*R*) isomer 5,273,995 5 6
The "trans racemate" of Scheme 1 means a mixture of 20 the following: -continued HO
25
30 Ph CONHPh I\ Ph CONHPb [R(R*R*)]isomer The conditions of the Step 1 and 2 of Scheme 1 are 35 generally in the Examples and 7 hereinafter. and as found 6 The chiral synthesis is shown in Scheme 2 (where Ph is phenyl) as follows:
Scheme 2 5,273,995 7 a -continued Scheme 2
0-
F
0
2. Tol. -H2O CO2Bu'
(6) 83% F
(6) 83% [a]d3= + 18.07 (CHCI3)
Generally, conditions for Scheme 2 are as shown in 60 the Exampies 1-5 hereinafter. which is now also incorporated by reference therefor. One of ordinary skill in the art would recognize van- The CSI data of the I, its the ations in the Schemes 1 and 2 which are appropriate for [I the racemateof these two oompounds of present the preparation the compounds of the inven- are as follows: tion. The compounds according to present invention and 65 especially according to the compound of the formula I ICs0 inhibit the biosynthesis of cholesterol as found in the Compound (micromoles/Liter) CSI screen that is disclosed in U.S. Pat. No. 4,681,893 [R-(R*R*)] isomer 0.0044 5,273,995 10 ice to -20' C. The light brown crystalline-product (Example 1A) is dried in vacuum oven at 40° C. The yield is 194 g. The product 1A is recrystallized from EtOAc at 5 -10' C. to yield 100 g to yield product 1B and then recrystallized from acetone/pentane to yield 90 g to yield product 1C. The mother liquor is combined Accordingly, the present invention is the pharmaceu- from tical composition prepared from the compound of the the wash of the crude material and recrystallized from EtOAc/Hexane. 33 g of shows the following: formula I or II or pharmaceutically acceptable salts 1B thereof. 10 HPLC: 97.42.17 of the R,S to S,S isomers. 28.5 g of 1C These compositions are prepared as described in U.S. shows the following: HPLC:95.7:3.7. The combined 1B Pat. No. 4,681,893 which is, therefore, again incorpo- and 1C is recrystallized from CHCl3 MeOH 10:1; pro- rated by reference here. viding a product 1F having a yield of 48.7 g of white Likewise, the present invention is a method of use as crystal. hypolipidemic or hypocholesterolemic agents. The l5 The mother liquor of the first aqueous wash is crys- compounds of the present invention utilized in the phar- talized (EtOAc/Heptane) to yield product 1D of 21.4 maceutical method of this invention are administered to g; HPLC: 71.56:25.52. the patient at dosage levels of from 10 to 500 mg per day The mother liquor of 1B and 1C is combined and which for a normal human adult of approximately 70 kg recrystallized from CHCl3/MeOH/Heptane to yield is a dosage of from 0.14 to 7.1 mg/kg of body weight 2o 55.7 g white crystals of product IG. per day. The dosages may be preferably from 0.5 to 1.0 1D is recrystallized from CHCl3/MeOH to yield the mg/kg per day. product 1H. The dosage is preferably administered as a unit dos- All mother liquor is combined,concentrated then the age form. The unit dosage form for oral or parenteral residue is dissolved in hot CHCl3/MeOH 10:1;put on a use may be varied or adjusted from 10 to 500 mg, pref- 25 silica gel column; rad eluted with EtOAc/Hexane erably from 20 to 100 mg according to the particular 40:60. The material crystallized out on the column and application and the potency of the active ingredient. the silica gel is extracted with CHCl3/MeOH and con- The compositions can, if desired, also contain other centrated. Recrystallization of the residue from active therapeutic agents. Determinations of optimum 3o CHCl3/Heptane 3:l yields 33.7 g of product II. dosages for a particular situation is within the skill of The mother liquor of IIis recrystallized to yield 18.7 the art. g of product 1K. The compounds of the formula I and II and their The mother liquor of 1K is crystallized to yield 6.3 g pharmaceutically acceptable salts are in general equiva- of product 1L. lent for the activity of the utility as described herein. 35 II, 1K and 1L is combined and recrystallized from The following examples illustrate particular methods CHC13/Heptane to yield 48 g. for preparing compounds in accordance with this in- The combined mother liquor of II, 1K, and 1L is vention. These examples are thus not to be read as limit- concentrated to yield 31 g of 1M. ing the scope of the invention. The product 1F provides the following data. EXAMPLE 1 40 285 ml 2.2M n-butyl lithium (in Hexane) is added Anal: IF dropwise to 92 ml diisopropylamine in 300 ml THF at m.p. 229-230' C. 50°-60° C. in a 1000 ml 1 neck flask via dropping funnel Calc. Found and under nitrogen. The well stirred yellow solution is c 77.84 77.14 allowed to warm to about -20' C.Then it is cannulated 45 H: 6.02 6.45 into a suspension of 99 g S(+)-2-acetoxy-1,1,2-tri- phenylethanol in 500 ml absolute THF, kept in a 2L-3 neck flask at -70' C. When addition is complete, the These data are consistent with the formula reaction mixture is allowed to warm to - 10° C. over a period two a 50 of hours. Meanwhile, suspension of 0.63 F mol MgBr2 is prepared by dropping 564 ml(0.63 mol) of bromine into a suspension of 15.3 g of magnesium (0.63 mol) in 500 ml THF plus in 3 L flask equipped with reflux condenser, and overhead stirrer. When this is completed, the MgBr2 suspension is cooled to -78' C. 55 and the enolate solution (dark brown) is cannulated into the suspension within 30 minutes. Stirring is continued for 60 minutes at -78' C. 150 g 5-(4-fluorophenyl)-2-(1- Ph methylethyl)-1-(3-oxopropyl)-N,4diphenyl- 1H-pyr- Ph role-3-carboxamide in 800 ml THF absolute was added 60 PhNHOC dropwise over 30 minutes; then stirred for 90 minutes at -78° C., then quenched with 200 ml AcOH at -78' C. This is removed to a cool bath, 500 ml of H2O is added and the mixture concentrated in vacuo at 40°-50° C. 500 EXAMPLE 2 ml of 1:1 EtOAc/Heptane is added to the yellowish 65 slurry and filtered. The filtrate is washed extensively 162 g (0.206M)of the combined products IF, lG, 1H with 0.5N HCI, then several times with H20 and finally and IL of Example 1 are suspended in 800 ml Me- with EtOAc/Heptane (3:1) that was cooled with dry thanol/THF (5:3).Cooled to 0" C.and added to 11.7 g 5,273,995 11 12 sodium methoxide. The mixture is stirred until every- Stirring is continued for four hours at -70' C. The thing is dissolved, then put in the freezer overnight. The reaction mixture is then quenched with 69 ml glacial reaction mixture is allowed to warm to room tempera- acetic acid and allowed to warm to room temperature. ture, quenched with 15 ml HOAc, then concentrated in The mixture is concentrated in and the is vacuo at C. to obtain expected product as follows: 5 40° taken up in EtOAc, washed with water extensively, then saturated NH4Cll, NaHCO3 (saturated), and finally with brine. The organic layer is dried aver anhydrous MgSO4, filtered and the solvent evaporated.The NMR 10 of the reaction is consistent with starting material plus product in about equal amounts plus some material on the baseline of the TLC. The material of the baseline of the TLC is separated from starting material and the . prduct is extracted by acid/base extraction. The or- 15 ganic phase is dried and concentrated in vacuo to yield 73 g. The NMR and TLC arc consistent with the for- mula
This product is added to 500 ml H2O and extracted 2o twice with EtOAc (300 ml). The combined extracts are washed with saturated NaHC03, brine, dried over an- hydrous magnesium sulfate, filtered and the solvent evaporated. The residue is chromatographed on silica (1:4) as 109 25 OH 0 gel in EtOAc/Heptane eluent to yield g Ph 0 colorless oil which is recrystallized from Et2O/Heptane II It to yield: OtBu 73.9 g first crop; white crystals PhNHCO 8.2 g second crop; white crystals. The crystals provide the following data: 30 m.p. 125°-126° C., ad20=4.23" (1.17M,CH3OH)
Calc. Found EXAMPLE 4 C: 72.76 12.5 I 35 73 g crude product of Example 3 is dissolved in 500 H: 6.30 6.23 N: 5.30 5.06 ml absolute THF and 120 ml triethyl borane is added, followed by 0.7 g t-butylcarboxylic acid. The mixture is stirred under a dry atmosphere for 10 minutes, cooled to These data are consistent with the formula -78' C. and 70 ml methanol is added and followed by 4.5 g sodium borohydride. The mixture is again stirred at -78' C. for six hours. Then poured slowly into a 4:l:l mixture of ice/30% H2O2/H2O This mixture is stirred overnight then allowed to warm to room tem- 45 perature. CHCl3 (400 ml) is added and the mixture is parti- tioned. The water layer is extracted again with CHCl3. The organic extracts are combined and washed exten- sively with H2O until no peroxide could be found. The PhNHCO 50 organic layer is dried over MgS04, filtered and the solvent is evaporated. The residue is treated by flash chromatography on silica gel, i.e. EtOAc/Hexane 1:3 to yield 51 g. The product is dissolved in THF/MeOH added EXAMPLE and 3 55 to 100 ml in NaOH, then stirred for four hours at room 77 ml of diisopropylamine is dissolved in 250 ml THF temperature. The solution is concentratad at room tem- in a ml three-neck flask equipped with thermome- perature to remove organic solvent, added to 100 ml ter and dropping funnel. The reaction mixture is kept H20, and extracted with Et20 twice, The aqueous layer under nitrogen. The mixture is cooled to -42° C. and is with HCl and extracted with EtOAc added to ml n-butyl lithium (in Hexane) 200 2.2M Of 60 three times, The combined layers are dropwise over 20 minutes and stirred for 20 minutes before adding dropwise 62 ml of t-butylacetate, dis- with H2O The organic layer is dried with anhydrous solved in 200 THF (over about 30 minutes).This MgSO4, filtered, and the solvent evaporated. The resi- mixture is stirred 30 minutes at -40° C., then 140 ml due is taken up in 2 liters of toluene and heated to reflux 2.2M of n-butyl lithium is added over 20 minutes. When 65 using a Dean-Stark trap for 10 minutes. addition is complete, 81 g of the product of Example 2 The reaction mixture is allowed to cool to room tem- in 500 ml absolute THE is added as quickly as possible perature overnight. Reflux is repeated for 10 minutes without allowing the temperature to rise above -40° C. and cooled for 24 hours. 5,273,995 13 14 The procedure above is repeated. The reaction is left [ß],[§]- dihydroxy-5-(1-methylethyl)-3-phenyl-4- at room temperature for the next 10 days, then concen- [(phenylamino carbonyl]-N-(1-1 -phenyleth y l- 1H-pyr- trated to yield 51 g of colorless foam. role-1-heptanamide, (hydroxy centers are both R) (1 g, This product is dissolved in minimum CHCl3 and 1.5 mmol) is added 1N NaOH (3.0ml, 3 mmol). The chromatographed on silica gel eluting with EtOAc/- 5 resulting solution is heated to reflux for 48 hours. Heptane (50:50) to yield 23 g in pure material. The solution is cooled to room temperature and con- Chromatography on silica gel in CHCl3/2-propanol centrated in vacuo. The residue is resuspended in water (98.5:1.5) yields 13.2 g. and carefully acidified with 6N HC1. The resulting acidic solution is extracted with ethyl acetate. The or- 10 Calc. ganic extract is washed with water, brine, dried over C: 73.31 MgS04, filtered and concentrated in vacuo. This resi- H: 615 due is redissolved in toluene (100 ml) and heated to N: 5.18 reflux with azeotropic removal of water for three hours. is cooled to room temperature and concentrated in 15 This vacuo to yield 1.2 g of a yellow semi-solid. Flash chro- EXAMPLE 5 matography on silica gel eluting with 40% EtOAc/- Preparation of 2R-trans-5-(4-fluorophenyl)-2-(1- Hexane gives 0.42 g of a white solid which still contanins methylethyl)-N,4-diphenyl- 1-[2-tetrahydro-4-hydroxy- impurities. This is rechromatographed to give 0.1 g of 6-oxo-2H-pyran-2-yl)ethyl]-1 H/-pyrrole-3-carboxa- 20 essentially pure R,R,enantiomer, 2R-trans-5-(4-fluoro- mide phenyl)-2-(1 -methylethyl)-N,4-diphenyl- 1 -[2-(-tetrahy- The product of Example 4 is recrystallized from dro-4-dydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1H-pyrrole- EtOAc/Hexane. Fraction 1 yields 8.20 g of 4A. The 3-carboxamide, as a white foam. HPLC shows this ma- mother liquor yields 4.60 g of 4B, HPLC of 4B shows terial to be 94.6% chemically pure in 100% of the product to be the [R*(R*R*)] isomer. 4A is 25 CHCl3=25.5°. The peak at room temperature=53.46 recrystallized to yield 4.81 g of 4C. 4B is chromato- minutes is tentatively assigned to an unknown diastereo- graphed silica gel CHCl3/2-propanol to yield 4.18 on in mer resulting from the 2% (S)-( -)-a-methylbenzyla- g colorless showing (0.53% foam of 4D ad23+24.53” in mine present in the Aldrich a-methylbenzylamine. CHCl3). 4C is recrystallized and the mother liquor of 4C is to yield 2.0 g.HPLC which indicates 100% of the EXAMPLE 8 R-trans isomer 2R-trans-5-(4-fluorophenyl)-2-(1- 30 methylethyl)-N,4-diphenyl- 1-[2-tetrahydro-4-hydroxy- Preparation of 2S-trans-5-(4-fluorophenyl)-2-(1- 6-oxo-2H-pyran-2-yl)ethyl]-1 H-pyrrole-3-carboxamide methylethyl)-N,4-diphenyl- 1-[2-(tetrahydro-4-Hydroxy- 6-oxo-2H-pyran-2-yl)ethyl]- lH-pyrroIe-3carboxamide- EXAMPLE 6 (S,S enantiomer of the compound prepared in Example Preparation of disastereomerica-methylbenzylamides 35 5 A solution of the racemate, trans-(+)-5-(4-fluoro- Carrying out the procedure described in Example 7 phenyl)-2(1 -methylethyl)-N,4-diphenyl-1-[2-tetrahy- on diastertomer 2 afforded 0.6 g of a foamy solid which dro-4-dydroxy-6-oxo-2H-pyran-2-yl)ethyl]-1 H-pyrrole- was flash chromatographed on silica gel. Elution with 3-carboxamide, (30 g, 55.5 ml) in (R)-(+)-a-methylben- 50% EtOAc/Hexane gave 0.46 g of essentially pure zylamine (575 ml, 4.45 mol, 98% Aldrich) is stirred 40 S,S, enantiomer 2S-trans-5-(4-fluorophenyl)-2-(1- overnight at room temperature. methylethyl)-N,4-diphenyl- 1-[2-(tetrahydro-4-hydroxy- The resulting solution is then diluted with ether (2 1) 6-oxo-2H-pryan-2-2yl)ethyl]- 1H-pyrrole-3-carboxamide, and then washed exhaustively with 2M HCI (4x500 as a white foam. HPLC showed this material to be ml), water (2 X 500 ml) and brine (2 x 500 ml). The or- 97.83% chemically pure. in ganic extract is then dried over MgSO4, filtered and 45 CHCl3= -24.8%. concentrated in vacuo to yield 28.2 g of the diastereom- eric a-methylbenzylamides as a white solid; m.p. EXAMPLE 9 174.0'-177°. The a-methylbenzylamides are separated Hydrolysis of chemical lactone of formula II by dissolving 1.5 g of the mixture in 1.5 ml of 98: 1.9:0.1 To a room temperature, solution of the lactone in CHCl3:CH3OH:NH4OH (1000 mg/ml and injecting 50 THF is added a solution of sodium hydroxide in water. onto a preparative HPLC column (silica gel, 300 The mixture is stirred for two hours HPLC:99.6% mmX41.4 mm I.D.) by gastight syringe and eluting (product); 0.34 to (starting lactone). The mixture is with the above solvent mixture. Fractions are collected diluted with 3 L water, extracted with ethyl acetate by UV monitor. Diastereomer 1 elutes at 41 minutes. (2~I L) and acidified to pHX4 by addition of 37 ml of Diastereomer 2 elutes at 49 minutes. Center cut frac- 55 5N hydrochloric acid. The aqueous layer is extracted tions are collected. This procedure is reputed three with 2X 1.5 L portions of ethyl acetate. The combined times and the like fractions are combined and concen- ethyl acetate extracts arc washed with 2x 1 L of water, trated. Examination of each by analytical HPLC indi- brine and dried, gave after filtration the ethyl acetate cates that diastereomer 1 is pure diastereo- 99.84% and solution of the required face-acid. This solution is used mer 2 is 96.53% pure. Each isomer is taken on sepa- directly in the fraction of the N-methylglucamine salt. rately to following Examples. 6o The ethyl acetate extracts from the brine-water were EXAMPLE 7 concentrated to give 15.5 g of an off-white solid. Preparation of 2R-trans-5-(4-fluorophenyl)-2-( 1- EXAMPLE 10 methylethyl)-N,4-diphenyl-1 -[2-(tetrahydro-4-hydroxy- 65 6-oxo-2H-pyran-2-yl)ethyl]- 1H-pyrrole-3-carboxamide Calcium Salt from Sodium Salt and/or Lactone To an ethanolic solution (5OM) of diastereomer 1 of Dissolve one mole lactone (540.6 g) in 5 L of MeOH; Example 6, [3R-[3R*(R*),5R*]]-2-(4-fluorophenyl)- after dissolution add 1 L H2O. While stirring, add one 5,273,995 15 16 equivalent NaOH and follow by HPLC until 2% or less -continued lactone and methyl ester of the diolacid remains (cannot use an EXCESS of NaOH, because Ca(OH)2 will form an addition of CaCl2). Charge NaOH as caustic (51.3 ml, 5 98 eq.) or as pellets (39.1 g, 0.98 eq). The above procedure is shown as follows:
10 0 m.w. = 1155.4 g
OH 15 EXAMPLE 11 98eq NaOH Treatment of Ethyl Acetate Solution of Free-acid of MeOH, H20 5:1 the Formula I With N-methylglucamine Ph To a solution of the free-acid of the formula I (0.106M) in ethyl acetate (3 L) is added a solution of N-methylglucamine (20.3 g, 0.106 m) in (1:l) water-ace- m.w = 540.6g tone (120 ml, 120 ml) with vigorous stirring at room temperature. Stirring is continued for 16 hours and the 1:1 OH OH 0 hazy solution concentrated in vacuo to ˜250 mp. Tolu- EtOAc, ene (1 L) is added and the mixture concentrated to a 25 white solid ˜100 g. The solid is dissolved in 1670 ml acetone and filtered into a three-neck flask equipped with a mechanical stirrer and thermostat controlled thermometer, The flask and filter is washed with 115 ml Ph (1:l) water-acetone and the clear solution is coaled 30 slowly. This provided a precipitate which is re-dis- solved by heating back to 65' C. Addition of a further Upon completion of hydrolysis, add 10 L H20, then 2o Of water by washing gives a line product which was isolated by filtration. The solids wash at least two times with a 1:1 mixture of EtOAc/- 35 washed with 1200 ml CH3CI and vacuum dried Hexane. Each wash should contain 10 L each of 255' to five a white solid. Analysis of this material EtOAc/Hexane. If sodium salt is pure, add 15 L of indicatesthat it contains 4% as well as 0.4% MeOH. If it is impure and/or contains color, add 100 residual acetone and 0.67% water. Analytical results are noted as follows: of G-60 charcoal, stir for two hours and filter over 4o Melting point: 105°-155°. C. Analysis supercel. Wash with 15 L MeOH. Perform a wt/vol % Expected: C=63.73; H-6.95; N=5.57: F2=9.53. Anal-
tion to 60° C. Add CaCl2 solution slowly, with high 256 nm: 1.0 ml/min. agitation. After complete addition, cool slowly to 15' C. 6-81 min.: 98.76% Opt. Ret.: [a].b= - 10.33' (c= 1.00, MeOH) adfilter. Wash filter cake with 5 L H20.Dry at 50° C. 5o Residual Solvents: CH2CH=0.26% in vacuum oven. Titrations: Can be recrystallized by dissolving in 4 L of EtOAc HClO4 (0.IN) =2O3.8% (50° C.) filtering over supercel, washing with 1 L Bu4NOH (O.1N)=98.5% Other salts prepared in a manner analogous to those EtOAc, then charging 3 L of hexane to the 50° C. rxn 55 processes appropriately selected from Examples 10 solution. 11 above may be the potassium salt, hemimagnesium The above procedure is shown as follows: salt, hemizine salt or the l-deoxy-2-(methylamino)-D- glucitol complex of the compound of formula I. I claim: OH OH 0 60 1. [R-(R*,R*)]-2-(4-fluoropheny)-ß,§-dihydroxy-5- (1 -methylethyl)-3-phenyl-4-[(phenylamino)-carbonyl]- IH-pyrrole-1-heptanoic acid or (2R-trans)-5-(4-fluoro- eq. CaCl2.2H20 phenyl)-2-( 1-methylethyl)-N,4-diphenyl- 1-1-[2-(tetrahy- H20 dro-4-hydroxy-6-oxo-2H-pyran-2-yl)ethyl]- 1H-pyrrole- Ph C 65 3-carboxamide; or pharmaccutically acccptable salts thereof. 2. A compound of claim 1 which is [R-(R*R*)]-2-(4- fluorophenyl)-ß-§6-dihydroxy-5-( 1-methyethyl)-3-phe- 5,273,995 17 18 nyl-4-[(phenylamino)carbonyl]-1H-pyrrole-1-heptanoic 8. The hemmimagnesium salt of the compound of claim acid. 2. 9. The hemizinc salt of the compound of claim 2. 3. A Of 1 which is (2R-trans-5-(4- 10. The l-deoxy-l-(methlamino)-D-glucitol fluorophenyl)-2-(1-methylethyl)-N,4-diphenyl-1- 1-[2-(tet- 5 with the of claim 2. rahydro-4-hydroxy-6-oxo-2H-pyran-3-yl)ethyl]- 1 H- 11. A pharmaceutical composition for treating hyper- pyrrole-3-carbxamide. cholesterolemia comprising a hypocholesterolrmicef- 4. The monosodium salt of the compound ofclaim 2. fective amount of a compound of claim 1 a Pharma- 5. The monopotassium salt of the compound of claim ceutically acceptable carrier. 12. A inhibiting cholesterol synthesis in a 2. 10 method of human suffering from hypercholesterolemiacomprising 6. The hemicalcium salt of the compound of claim 2. administering compound of dosage 7. The N-methylglucamine salt of the compound of form. claim 2. IS
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65 07 660976 PATENT APPLICATION SERIAL NO.
U.S. DEPARTMENT OF COMMERCE PATENT AND TRADEMARK OFFICE FEE RECORD SHEET
PTO- 15 5 6 5/87 PATENT/DOCKET NO: 3844-01-JT
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JULY 21, 1989 ELIZABETH M. ANDERSON AGENT Date Registration No.31,585 Warner-Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 Tel. NO. (313) 996-7304
Attachment: Triplicate Copies for Deposit Account Authorization
PLD-51A Page 2 of 2 2/89 -24-
.
ABSTRACT
acid or 5 and pharmaceutically acceptable salts thereof. .wl PD-3844-01-JT
METHYLETHYL-3-PHENYL-4- CARBONYL] ACID, ITS LACTONE FORM AND SALTS THEREOF ACID, ITS LACTONE FORM AND SALTS THEREOF
, , 5 BACKGROUND OF THE INVENTION
are among compounds of 10 U.S. Patent No. 4,681,893 having usefulness as inhibitors of cholesterol biosynthesis. The compounds therein broadly include 4-hydroxypyran-2-ones and the corresponding ring-opened acids derived therefrom. It is now unexpectedly found that the enantiomer having 15 the R form of the ring-opened acid of
I
pyrrole-3-carboxamide; that
20 acid, provides surprising inhibition of the biosynthesis of cholesterol. It is known that 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) exists as the 3R-stereoisomer. Additionally, as 25 shown in the study of a series of 5-substituted acids by Stokker et al,, in A Reductase Inhibitors. 1. Structural Modification of 5-Substituted acids and Their Lactone Derivatives," i J. Med. Chem. 1985, 28, 347-358, essentially all of the biological activity resided in the trans diastereomer of
hydroxy-2H-pyranone having a positive rotation. Further, . the absolute configuration for the 35 moiety common to mevinolin of the formula (la) -2-
. .
la
and compactin of the formula (lb)
1b
H
apparently is required for inhibition of HMG-CoA reductase. 5 This is reported by Lynch et al. in "Synthesis of an HMG-CoA Reductase Inhibitor; A Diastereoselective Aldol Approach in Tetrahedron Letters, Vol. 28, No. 13, pp. 1385-1388 (1987) as the 4R, 6R configuration. However, an ordinarily skilled artisan may not predict 10 the unexpected and surprising inhibition of cholesterol biosynthesis of the present invention in view of these disclosures. SUMMARY OF THE
Accordingly the present invention provides for compounds consisting of
5 acid (compound of formula I), pharmaceutically acceptable salts thereof and
(the lactone form of the 10 heptanoic acid or cornpound of formula The present invention also relates to a pharmaceutical composition, useful as a hypocholesterolemic agent, comprising a hypocholesterolemic effective amount of
. 15 acid, its pharmaceutically acceptable salts or
pyrrole-3-carboxamide acid; and a pharmaceutically 20 acceptable carrier. Further, the present invention is also a method of treating mammals, including humans, suffering from hypercholesterolemia by administering to such mammal a dosage form of the pharmaceutical composition described above.
25 DETAILED DESCRIPTION OF THE INVENTION
The pharmaceutically acceptable salts of the invention are those generally derived by dissolving the free acid or the lactone; preferably the lactone, in aqueous OK aqueous alcohol solvent or other suitable solvents with an appro- 30 priate base and isolating the salt by evaporating the solution or by reacting the free acid or lactone; preferably the lactone and base in an organic solvent in which the salt separates directly or can be obtained by concentration of the solution. -4-
In practice, use of the salt form amounts to use of the acid or lactone form. Appropriate pharmaceutically accept- able salts within the scope of the invention are those derived from bases such as sodium hydroxide, potassium 5 hydroxide, lithium hydroxide, calcium hydroxide, 1-deoxy-2- magnesium hydroxide, zinc hydroxide, aluminum hydroxide, ferrous or ferric hydroxide, ammonium hydroxide or organic amines such as N-methylglucamine, choline, arginine and the like. 10 Preferably, the lithium, calcium, magnesium, aluminum and ferrous or ferric salts are prepared from the sodium or potassium salt by adding the appropriate reagent to a solution of the sodium or potassium salt, i.e., addition of calcium chloride to a solution of the sodium or potassium 15 salt of the compound of the formula I will give the calcium salt thereof. The free acid can be prepared by hydrolysis of the lactone form of formula II or by passing the salt through a cationic exchange resin (H + resin) and evaporating the 20 water. The most preferred embodiment of the present invention is
1-heptanoic acid, hemicalcium salt. 25 Generally, the compounds I and II of the present invention can be prepared by (1) resolving the racemate, that is prepared by the processes described in U.S. Patent No. 4,681,893 which is incorporated by reference therefor, or (2) synthesizing the desired chiral form beginning from 30 starting materials which are known or readily prepared using processes analogous to those which are known. Specifically, resolution of the racemate may be accomplished as shown in Scheme 1 (where Ph is phenyl) as follows -5- .;-".;,.--
t
Step A t F trans racemate
Step B 1) Separate 2) NaOH 3) reflux in toluene
e
. . -6- . The "trans racemate" of Scheme 1 means a mixture of the following:
Ph CONHPh Ph CONHPh
[R(R*R*) isomer (R*R*) isomer
-..----
The conditions of the Step 1 and 2 of Scheme 1 are 5 generally as found in the Examples 6 and 7 hereinafter. -7-
The chiral synthesis is shown in Scheme 2 (where Ph is phenyl) as follows:
Scheme 2
1. lhr 2. AcOH
F
0 1.1 eq NaOMe MeOH, -10°C 16hrs Ph PhOH PhNHOC
73% (3) 75% (4
1. , NaBH4
-3O--40°C 5hrs
F t
OH OH 1. NaOH 2. Tol.
(6) 83%
, -8-
i Generally, conditions for Scheme 2 are as shown in the Examples 1-5 hereinafter, One of ordinary skill in the art would recognize variations in the Schemes 1 and 2 which are appropriate for 5 the preparation of the compounds of the present invention, The compounds according to present invention and especially according to the compound of the formula I inhibit the biosynthesis of cholesterol as found in the CSI screen that is disclosed in U.S. Patent No. 4,681,893 which 10 is now also incorporated by reference therefor. The CSI data of the compound I, its enantiomer the compound II and the racemate of these two compounds are as follows: -- -
Compound (micromoles/liter) 15 [R-(R*R*)] isomer 0.0044 [S-(R*R*)J isomer 0.44 Racemate 0.045
Accordingly, the present invention is the pharmaceu- tical composition prepared from the compound of the 20 formula I or II or pharmaceutically acceptable salts thereof. These compositions are prepared as described in U.S. F Patent No. 4,681,893 which is, therefore, again incorporated by reference here. 25 Likewise, the present invention is a method of use as hypolipidemic or hypocholesterolemic agents. The compounds of the present invention utilized in the pharmaceutical method of this invention are administered to the patient at dosage levels of from 10 to 500 mg per day which for a 30 normal human adult of approximately 70 kg is a dosage of from 0.14 to 7.1 mg/kg of body weight per day. The dosages may be preferably from 0.5 to 1.0 mg/kg per day. The dosage is preferably administered as a unit dosage form. The unit dosage form for oral or parenteral use may 35 be varied or adjusted from 10 to 500 mg, preferably from 20 to 100 mg according to the particular application and the -9-
potency of the active ingredient. The compositions can, if desired, also contain other active therapeutic agents. Determinations of optimum dosages for a particular situation is within the skill of the art. 5 The compounds of the formula I and II and their pharma- ceutically acceptable salts are in general equivalent for the activity of the utility as described herein. The following examples illustrate particular methods for preparing compounds in accordance with this invention. These examples are thus not to be read as limiting the scope of the invention.
EXAMPLE 1 285 ml 2.2 M n-butyl lithium (in Hexane) is added dropwise to 92 ml diisopropylamine in 300 ml THF at 50-60°C in a 1000 ml 1 neck flask via dropping funnel and under nitrogen. The well stirred yellow solution is allowed to warm to about -20°C. Then it is cannulated into a suspension of 99 g in 500 ml absolute THF, kept in a 2L-3 neck flask at -70°C. When addition is complete, the reaction mixture is allowed to warm to -100C over a period of two hours. Meanwhile, a suspension of 0.63 mol MgBr2 is prepared by dropping 564 ml (0.63 mol) of bromine into a suspension of 15.3 g of magnesium (0.63 mol) in 500 ml THF plus in 3L flask equipped with reflux condenser, and overhead stirrer. When this is completed, the MgBr2 suspension is cooled to -78°C and the enolate solution (dark brown) is cannulated into the suspension within 30 minutes. Stirring is continued for 60 minutes at -78°C. 150 g
carboxamide in 800 ml THF absolute was added dropwise over 30 minutes; then stirred for 90 minutes at -78°C, then quenched with 200 ml AcOH at -78°C. This is removed to a cool bath, 500 ml of H20 is added and the mixture 35 concentrated in vacuo at 4O-5O°C. 500 ml of 1:1 -10-
EtOAc/Heptane is added to the yellowish slurry and fi tered The filtrate is washed extensively with 0.5 N Hc1, then several times with H20 and finally with EtOAc/Heptane (3:l) that was cooled with dry ice to -2O°C. The light brown crystalline product (Example 1A) is dried in vacuum oven at 4O°C. The yield is 194 g. The product 1A is recrystallized from EtOAc at -10°C to yield 100 g to yield product 1B and then recrystallized from acetone/pentane to yield 90 g to yield product 1C. The 10 mother liquor is combined from the wash of the crude material and recrystallized from EtOAc/Hexane. 33 g of lB shows the following: HPLC: 97.4:2.17 of the R,S to S,S isomers. 28.5 g of 1C shows the following: HPLC:95.7:3.7. The combined 1B and 1C is recrystallized from CHCl 3 MeOH 15 10:l; providing a product IF having a yield of 48.7 g of white crystal. The mother liquor of the first aqueous wash is crystallized (EtOAc/Heptane) to yield product 1D of 21.4 g; HPLC: 71.56:25.52. 20 The mother liquor of 1B and 1C is combined and recrystallized from CHC13/MeOH/Heptane to yield 55.7 g white crystals of product 1G. 1D is recrystallized from CHC13/MeOH to yield the product 1H. All mother liquor is combined, concentrated then the residue is dissolved in hot CHC13/MeOH 10:l; put on a silica gel column; and eluted with EtOAc/Hexane 40:60, The material crystallized out on the column and the silica gel is extracted with CHC13/MeOH and concentrated. 30 Recrystallization of the residue from CHC13/Heptane 3:l yields 33.7 g of product 11. The mother liquor of 1I is recrystallized to yield 18.7 g of product 1K. The mother liquor of 1K is crystallized to yield 6.3 g 35 of product 1L. 11, 1K and 1L is combined and recrystallized from CHC13/Heptane to yield 48 g. -11-
The combined mother liquor of 11, lK, and 1L is concentrated to yield 31 g of 1M. The product 1F provides the following data.
Anal: 1F 5 m.p. 229-23O°c Calc. Found C: 77.04 77.14 H: 6.02 6.45 N: 3.56 3.13
10 These data are consistent with the formula
F
Ph Ph
Ph PhNHCO
EXAMPLE 2 162 g (0.206 M) of the combined products lF, 1G, 1H and 1 L of Example 1 are suspended in 800 ml Methanol/THF (5:3). 15 Cooled to 0°C and added to 11.7 g sodium methoxide, The mixture is stirred until everything is dissolved, then put in the freezer overnight. The reaction mixture is allowed to warm to room temperature, quenched with 15 ml HOAc, then concentrated in vacuo at 4O°C to obtain expected product as 20 follows: Ph 0 PhNHC
This product is added to 500 ml H20 and extracted twice with EtOAc (300 ml). The combined extracts are washed with saturated brine, dried over anhydrous magnesium 5 sulfate, filtered and the solvent evaporated. The residue is chromatographed on silica gel in as eluent to yield 109 g colorless oil which is recrystallized from to yield: 73.9 g first crop; white crystals 10 8.2 g second crop; white crystals. The crystals provide the following data:
4.23' (1.17 M,
Calc Found C: 72.76 72.51 15 H: 6.30 6.23 N: 5.30 5.06
These data are consistent with the formula -13-
. . .. .
. , . .
! .
EXAMPLE 3 77 ml of diisopropylamine is dissolved in 250 ml THF in a 2000 ml three-neck flask equipped with thermometer and 5 dropping funnel. The reaction mixture is kept under nitrogen. The mixture is cooled to -42°C and added to 200 ml 2.2 M of n-butyl lithium (in Hexane) dropwise over 20 minutes and stirred for 20 minutes before adding dropwise 62 ml of t-butylacetate, dissolved in 200 ml THF (over about 10 30 minutes). This mixture is stirred 30 minutes at -4O°C, then 140 ml 2.2 M of n-butyl lithium is added over 20 minutes. When addition is complete, 81 g of the product of Example 2 in 500 ml absolute THF is added as quickly as possible without allowing the temperature to rise above 15 -4O°C. Stirring is continued for four hours at -7O°C. The reaction mixture is then quenched with 69 ml glacial acetic acid and allowed to warm to room temperature. The mixture is concentrated in vacuo and the residue is taken up in EtOAc, washed with water extensively, then saturated NH4C1, 20 NaHC03 (saturated), and finally with brine. The organic layer is dried over anhydrous MgSO4, filtered and the solvent evaporated. The NMR of the reaction is consistent with starting material plus product in about equal amounts plus some material on the baseline of the TLC. The material 25 of the baseline of the TLC is separated from starting material and the product is extracted by acid/base -14-
extraction. The organic phase is dried and concentrated in vacuo to yield 73 g. The NMR and TLC are consistent with the formula
F
5 EXAMPLE 4 73 g crude product of Example 3 is dissolved in 500 ml absolute THF and 120 ml triethyl borane is added, followed by 0.7 g t-butylcarboxylic acid. The mixture is stirred under a dry atmosphere for 10 minutes, cooled to -78°C and 10 70 ml methanol is added and followed by 4.5 g sodium borohydride, The mixture is again stirred at -78°C for six hours. Then poured slowly into a 4:1:1 mixture of ice/30% H2O2/H2O. This mixture is stirred overnight then allowed to warm to room temperature. 15 CHCl3 (400 ml) is added and the mixture is partitioned. The water layer is extracted again with CHCl3. The organic extracts are combined and washed extensively with H20 until no peroxide could be found. The organic layer is dried over MgS04, filtered and the solvent is evaporated. 20 The residue is treated by flash chromatography on silica gel, i.e. EtOAc/Hexane 1:3 to yield 51 g. The product is dissolved in THF/MeOH and added to 100 ml in NaOH, then stirred for four hours at room temperature. The solution is concentrated at room 25 temperature to remove organic solvent, added to 100 ml H20, -15-
and extracted with twice. The aqueous layer is acidified with 1 N HC1 and extracted with EtOAc three times. The combined organic layers are washed with The organic layer is dried with anhydrous filtered, and the solvent evaporated. The residue is taken up in 2 liters of toluene and heated to reflux using a Dean-Stark trap for 10 minutes. The reaction mixture is allowed to cool to room temperature overnight. Reflux is repeated for 10 minutes and cooled for 24 hours. The procedure above is repeated. The reaction is left at room temperature for the next 10 days, then concentrated to yield 51 g of colorless foam. This product is dissolved in minimum and chromatographed on silica gel eluting with to yield 23 g in pure material. Chromatography on silica gel in yields 13.2 g.
Calc.
20 C: 73.31 H: 6.15 N: 5.18
EXAMPLE 5 Preparation of 25
The product of Example 4 is recrystallized from EtOAc/Hexane. Fraction 1 yields 8.20 g of 4A. The mother liquor yields 4.60 g of 4B, HPLC of 4B shows 100% of the 30 product to be the isomer. 4A is recrystallized to yield 4.81 g of 4C. 4B is chromatographed on silica gel in to yield 4.18 g colorless foam of 4D showing + (0.53% in . 4C is recrystallized -16-
and the mother liquor of 4C is to yield which indicates 100% of the R-trans isomer
5 pyrrole-3-carboxamide.
EXAMPLE 6 Preparation of diastereomeric , A solution of the racemate,
10 (30 g, 55.5 ml) in (575 ml, 4.45 mol, 98% Aldrich) is stirred overnight at room temperature. The resulting solution is then diluted with ether (2 1) , 15 and then washed exhaustively with 2 M HC1 (4 x 500 ml), water (2 x 500 ml) and brine (2 x 500 ml). The organic extract is then dried over filtered and concentrated in vacuo to yield 28.2 g of the diastereomeric as a white solid; The 20 a-methylbenzylamides are separated by dissolving 1.5 g of the mixture in 1.5 ml of (1000 and injecting onto a preparative HPLC column (silica gel, 300 mm X 41.4 mm I.D.) by gastight syringe and eluting with the above solvent mixture. Fractions are 25 collected by monitor. Diastereomer 1 elutes at 41 minutes. Diastereomer 2 elutes at 49 minutes. Center cut fractions are collected. This procedure is repeated three times and the like fractions are combined and concentrated. Examination of each by analytical HPLC 30 indicates that diastereomer 1 is 99.84% pure and diastereomer 2 is 96.53% pure. Each isomer is taken on separately to following Examples. -17-
EXAMPLE 7 Preparation of
5 To an ethanolic solution (50 M) of diastereomer 1 of , Example 6,
(hydroxy centers are both R) (1 g, 1.5 mol) is added 10 1 N NaOH (3.0 ml, 3 nmol). The resulting solution is heated to reflux for 48 hours. The solution is cooled to room temperature and concentrated in vacuo. The residue is resuspended in water and carefully acidified with 6 N HC1. The resulting acidic 15 solution is extracted with ethyl acetate. The organic extract is washed with water, brine, dried over MgS04, filtered and concentrated in vacuo. This residue is redissolved in toluene (100 ml) and heated to reflux with azeotropic removal of water for three hours. This is cooled 20 to room temperature and concentrated in vacuo to yield 1.2 g of a yellow semi-solid. Flash chromatography on silica gel eluting with 40% EtOAc/Hexane gives 0.42 g of a white solid which still contains impurities. This is rechromatographed to give 0.1 g of essentially pure R,R, enantiomer, 2R-trans- 25
as a white foam. HPLC shows this material to be 94.6% chemically pure in = 25.5°. The peak at room temperature = 53.46 minutes is tentatively 30 assigned to an unknown diastereomer resulting from the 2% present in the Aldrich a-methylbenzylamine. -18-
EXAMPLE 8 Preparation of 0
5 (S,S enantiomer of the compound prepared in Example 5 Carrying out the procedure described in Example 7 on diastereomer 2 afforded 0.6 g of a foamy solid which was flash chromatographed on silica gel. Elution with 50% gave 0.46 g of essentially pure S,S, enantiomer 10
as a white foam. HPLC showed this material to be 97.83% chemically pure. = 0.51% in CHC13 -24.8%.
1 15 EXAMPLE 9 Hydrolysis of chemical lactone of formula II To a room temperature, solution of the lactone in THF is added a solution of sodium hydroxide in water. The mixture is stirred for two hours (product); 0.34 20 to (starting lactone). The mixture is diluted with 3L water, extracted with ethyl acetate (2 X 1L) and acidified to pH X 4 by addition of 37 ml of 5N hydrochloric acid. The aqueous layer is extracted with 2 X 1.5L portions of ethyl acetate. The combined ethyl acetate extracts are washed 25 with 2 X 1L of water, brine and dried, gave after filtration the ethyl acetate solution of the required face-acid. This solution is used directly in the fraction of the N-methylglucamine salt. The ethyl acetate extracts from the brine-water were 30 concentrated to give 15.5 g of an off-white solid. -19-
EXAMPLE 10 Calcium Salt from Sodium Salt and/or Lactone I Dissolve one mole lactone (540.6 g) in 5 L of MeOH; after dissolution add 1L H2O. While stirring, add one 5 equivalent NaOH and follow by HPLC until 2% or less lactone and methyl ester of the diolacid remains (cannot use an excess of NaOH, because Ca(OH)2 will form an addition of CaCl2). Charge NaOH as caustic (51.3 ml, 98 eq.) or as pellets (39.1 g, .98 eq.).
10 The above procedure is shown as follows:
----- a
Ph c .98 eq. NaOH 0 N H Ph MeOH, H20 5:1 m.w.= 540.6 g
1:1 EtOAc, Hexane H20 F
Wash
Upon completion of hydrolysis, add 10 L H2O, then wash at least two times with a 1:1 mixture of EtOAc/Hexane. Each wash should contain 10 L each of EtOAc/Hexane. If sodium 15 salt is pure, add 15 L of MeOH. If it is impure and/or contains color, add 100 g of G-60 charcoal, stir for two hours and filter over supercel. Wash with 15 L MeOH. Perform a wt/vol % on the reaction mixture, by HPLC, to determine the exact amount of salt in solution. 20 Dissolve 1 eq. or slight excess CaC12=2H20 (73.5 g) in 20 L H2O. Heat both reaction mixture and CaC12 solution to 60°C. Add CaCI2 solution slowly, with high agitation. -20-
After complete addition, cool slowly to 15°C and filter. Wash filter cake with 5 L H2O. Dry at 50°C in vacuum oven. Can be recrystallized by dissolving in 4 L of EtOAc (50°C)filtering over supercel, washing with 1 L EtOAc, then 5 charging 3 L of hexane to the 5O°C rxn solution. The above procedure is shown as follows:
OH OH 0
1/2 eq.CaC12 2H20 ,
m.w.= 580.6 g
.
EXAMPLE 11 Treatment of Ethyl Acetate Solution of Free-acid of the 10 Formula I with N-methylqlucamine To a solution of the free-acid of the formula I (0.106 M) in ethyl acetate (3 L) is added a solution of N-methylglucamine (20.3 g, 0.106 m) in (1:1) water-acetone (120 ml, 120 ml) with vigorous stirring at room temperature. 15 Stirring is continued for 16 hours and the hazy solution concentrated in vacuo to - 250 mp. Toluene (1 L) is added and the mixture concentrated to a white solid - 100 g. The solid is dissolved in 1670 ml acetone and filtered into a three-neck flask equipped with a mechanical stirrer and 20 thermostat controlled thermometer. The flask and filter is washed with 115 ml (1:1)water-acetone and the clear solution is cooled slowly. This provided a precipitate which is re-dissolved by heating back to 65°C. Addition of a further 20 ml of water followed by the washing gives a
I . .- -21-
crystalline product which was isolated by filtration. The solids are washed with 1200 ml CH3C1 and vacuum dried at 255° to give a white solid. Analysis of this material indicates that it contains 4% amine as well as 0.4% residual 5 acetone and 0.67% water. Analytical results are noted as follows : Melting point: 105-155°C (decomposition) Analysis Expected: C = 63.73; H - 6.95; N = 5.57; F2 = 9.53. 10 Analysis Found: C 62.10; H 6.89; N - 5.34; F2. C 61.92; H - 7.02; N 5.38; F2. H2O = 0.47% (KF) HPLC: MeOH, H20, THF (40; 550; 250) Econosil: C18, 5µ , 25 CM 15 256 nm: 1.0 ml/min. . 6-81 min.: 98.76% \ Opt. Ret,: [a].b = -10.33' (c = 1.00, MeOH) Residual Solvents: CH2CH = 0.26% Titrations: HC104 (0.1 N) = 203.8% 20 Bu4NOH (0.1 N) = 98.5%
Other salts prepared in a manner analogous to those processes appropriately selected from Examples 10 and 11 above may be the potassium salt, hemimagnesium salt, hemizinc salt or the 25 complex of the compound of formula I. -22-
CLAIMS
1,
pyrrole-1-heptanoic acid or (2R-trans) fluorophenyl) -2- 1-methylethyl-N, 2- 5 and pharmaceutically acceptable salts thereof.
2. A compound of Claim 1
heptanoic acid.
3. A compound of Claim 1 which is
4. The monosodium salt of the compound of claim
5. The monopotassium salt of the compound of claim
6. The hemicalcium salt of the compound of claim
7. The N-methylglucamine salt of the compound of claim
8. The hemimagnesium salt of the compound of Claims
9. The hemizinc salt of the compound of claim
10. The mixture with the compound of claim -23-
11. A pharmaceutical composition for treating hypercholesterolemia comprising a effective amount of a compound pharmaceutically acceptable carrier.
12. A method of inhibiting cholesterol synthesis in a human \ suffering from administering a form.
I PAT/
PATENT/Docket No.: 3844-01-JT DECLARATION (37 CFR 1.63) AND POWER OF ATTORNEY a below-named inventor, I hereby declare that: My residence, post office address, and citizenship are as stated below next to my name and I I believe I am the original, first, and sole inventor (if only one name is listed below) original, first, and joint , inventor (if plural names are listed below) of the subject matter which is claimed and for which a patent is sought on the, invention entitled:
I ACID, ITS LACTONE FORM i AND SALTS THEREOF the specification of which is attached hereto.
was filed on as Application Serial No. hereby state that I have reviewed and understand the contents of the above-identified specification, including the claims. acknowledge the duty to disclose information which is material to the examination of this application in accordance with 37 CFR 1.56 (a). I I hereby declare that all statements made herein of my own knowledge are true and that all statements made on information and belief are believed to be true; and further that these statements were made with the knowledge that willful false statements and the like so made are punishable by fine or imprisonment, or both, under Section 1001 of Title 18 of the United States Code and that such willful false statements may jeopardize the validity of the application any patent issued thereon. I hereby appoint
Elizabeth M. Anderson Reg. No. 31,585) Craig M. Bell ...... (Reg. No 812 ) Ronald A. Daignault .... Reg. No. 25,968) Geoffrey G. Dellenbaugh Reg. No. 26,864) Sandra Field Reg. No. 32,568) Charles Gaglia Reg. NO. 31,014) Ruth H. Newtson Reg. No. 26,657) Daniel A. Scola Reg. NO Joan Thierstein Reg. No. 29,450) Francis J. Tinney Reg. No. 33,069 . . registered to practice before the Patent and Trademark Office as my attorneys/agent with full power of substitution and revocation to prosecute this application and all divisions and
Form Page 1 7/86 of 2 PATENT/Docket NO.: 3844-01- ,I continuations thereof and to transact all business in the Patent and Trademark Off ice connected therewith and request that all correspondence and telephone communications be directed to the following person(s) at the mailing address and telephone number hereafter given: I Name Joan Thierstein Registration No.: 29,450
Address : Patent Department Warner-Lambert Company 2800 Plymouth Road Ann Arbor, Michigan 48105
Telephone No. : (313) 996-7190
Full name of sole or first inventor /- Inventor's Signature
Residence 6089 White Swan Lane I post Office Address Ann Arbor, ,Michigan 48108
Citizenship United States of America
Residence Post Office Address Citizenship Full name of third joint inventor, if any
Residence i I I
PLD-63.2a/Short Form Page 2 7/86 of 2 PATENT/Doc, .
of mailing 37 CFR 1.10) date of deposit July 21, 1989 I hereby certify that this transmittal together with the. patent application referred to below is being daposited with the united States Postal Service express mail Post office to Addressee" service under 37 CFR 1.10 on the date idicated above and is addressed to the Commissioner of patents adtrademarks, washington D.C. 20231.
Penny Drake name of Person Hailing Paper
COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231 Sir:
Transmitted herewith for filing is the complete patent application of:
InVentor(s): Bruce David Roth
For: ACID, ITS LACTONE FORM AND SALTS THEREOF Additionally enclosed are:
sheets of drawing(s). Associate Power of Attorney. An assignment of the invention to A certified copy of a application. This application is being mailed by Express Mail under 37 CFR 1.10 and the required certificate appears above.
The filing fee has been calculated as shown below:
Basic Fee For number Filed number Extra Rate $370.00
Total Claims 12 - 20 -0- x $12 -0-
Independent Claims 1- 3 -0- x 536 -0-
1 Recording Fee for Assignment $8.00
multiple Dependent Claim presented x $120
TOTAL filing FEE $370.00
PLD-51A Page 1 of 2 2/89 t - PATENT/DoC NO. 3844-01-JT
art Unit Examiner
APPlicant(s) : BRUCE DAVID ROTH Serial Number : Filed For:
ACID, ITS LACTONE FORM AND SALTS THEREOF Commissioner of Patents and Trademarks Washington, D.C. 20231 INFORMATION DISCLOSURE STATEMENT UNDER 37 CFR 1.97 AND 1.98 Sir: Provided herewith are copies of patents and publica- tions as listed in the attached form A concise explanation of the relevance of each reference listed in Form PTO-1449 is: [X] contained in the above-captioned specification as filed; and/or provided in a communication enclosed with this paper. Applicant respectfully asserts that the substantive provisions of 37 CFR 1.97 and 37 CFR 1.98 are met by the provision of the foregoing statement.
Respectfully submitted,
JULY 21, 1989 ELIZABETH M. ANDERSON , AGENT Date Registration N0.31,585 Warner-Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 Tel. No. (313)996-7304
Attachment: PTO-1449
PLD-97.1 11/87 atty.docketF PD-38 4 4-0 T applicant BRUCE D. ROTH
FOREIGN PATENT DOCUMENTS translation documentnumber date country
examiner
EXAMINER: initial it citation considered whether or not citation is in conformance with MPEP 609 draw line through citation if not in conformance and not considered. Include copy of this farm with next communication to applicant UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office Address : COMMISSIONER OF PATENTS AND TRADEMARKS Washington. D.C 20231
3
This application has been examined Responslve to communication flied on This action is made final.
A shortened statutory period for response to thls action Is set to expire days from the date of this letter. Failure to respond within the period for response will cause the application to become abandoned. 35 U.S.C. 133
Part I THE FOLLOWING ATTACHMENT(s) ARE PART OF THIS ACTION:
1. Notice of References Cited by Examiner, PTO-892. 2. Notlce re Patent Drawing, PTO-948. 3. Notice of Art Cited by Applicant, PTO-1449. 4. Notice of informal Patent Application, Form PTO-152. 5. information on How to Effect Drawing Changes, PTO-1474. 6.
Part II SUMMARY OF ACTION
Claims are pending in the application.
Of the above. claims are withdrawn from consideration.
2. Claims have been cancelled.
3. Claims are allowed.
4. Claims are rejected.
5. Claims are objected to.
Claims are subject to restriction or election requirement,
7. This application has been filed with lntormal drawings under 37 C.F.R. 1.65 which are acceptable for examination purposes.
8. Formal drawings are required In response to this Office action.
9. The corrected or substitute drawings have been received on . Under 37 C.F.R. 1.84 these drawings are acceptable. not acceptable (see explanation or Notice re Patent Drawing, PTO-948).
10. The proposed additional or substitute sheet(s) of drawings, filed on has (have) been approved by the examiner. disapproved by the examiner (see explanation).
11. The proposed drawing correction, filed on , has been approved. disapproved (see explanation).
12. Acknowledgment is made of the claim for priority under U.S.C. 119. The certified copy has been received not been received
been filed In parent application, serial no. filed on
13. Since this application appears to be In condition for allowance except for formal matters, prosecution as to the merits is closed In accordance with the practice under Ex parte Quayle. 1935 C.D. 11: 453 O.G.213.
14. Other
EXAMINER'S ACTION PTOL-326 (Rev. 6-88) Serial No. 07/384187 -2- Art Unit 121
Claims 1 to 12 are currently pending in the
application. All cited references have been considered, and any references cited but not used in a rejection are cited to show the state of the art.
Claims 1, 2 and 3 are rejected under 35 U.S.C. 112, second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which applicant reqards as the invention.
In the first-named compound of claim 1, and in claim 2, there are two parentheses before "1-methylethyl", but only one closing parentheses making the name difficult to translate into a structural
formula. Similarly, in the second-named compound of
claim 1 and in claim 3, there is a beginning parentheses
before but no closing parentheses. The following a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless- (b) the invention was patented or described in a Printed publication in this or a foreign country or in public use or on sale in this country, more than one (1) year prior to the date of application for patent in the United States. Claims 1 to 12 are rejected under 35 U.S.C. 102 (b)
AS being anticipated by Roth, U.S. Patent 4,681,893. Roth teaches the claimed substituted alkyl 1 2-ones and their corresponding dihydroxy -acids as useful inhibitors of cholesterol Serial No. 071384187 -3 -
Art Unit 121
synthesis. See column 2, formula I and lines 39-43: column 6 formula X. Reqarding the stereochemistry of these compounds, column 3 Lines 49 to 54 restrict the invention to the trans-isomers and column 6 lines 56-58 specify the R*, R* configuration. Thus, the claimed compounds, salts, compositions, and method are considered to be anticipated by Roth. Any inquiry concerning this communication or earlier communications from the examiner should be directed to Examiner Mary Sue Howard whose telephone number is (703) 557-1213.
any inquiry of a general nature, or relating to the status of this application, should be directed to the
Group receptionist whose telephone number is (703)
557-3920.
03/19/90:dal August 6, 1990 1 together with the reply and/or amendment referred to below is being deposited with the United States Postal Service as first class mail under 37 CFR 1.8 on the date indicated above and is addressed to the Commissioner of Patents and Trademarks, Washington D.C. 20231,
Penny Drake Name of Person Mailing Paper Signature
IN THE UNITED STATES PATENT AND TRADEMARK OFFICE !
Art Unit 121 Examiner M. Howard Applicant(s) Bruce David Roth Serial Number 07/384,187 '3 Filed July 21, 1989 For [R- (R*R*) -2-- (1-
ACID, ITS LACTONE FORM AND SALTS Commissioner of Patents and Trademarks THEREOF Washington, D.C. 20231
TRANSMITTAL OF A RESPONSE TO A NON-FINAL ACTION (37 CFR 1.111)
Sir:
Transmitted herewith Is a reply and/or amendment in the above-captioned application in response to the Examiner's action dated March 22, 1990
The reply and/or amendment is being filed under 37 CFR 1.8 and the required Certificate of Mailing appears above. The use of this certificate is intended to insure that the reply and/or amendment will be considered as timely filed.
An additional fee in the mount of $ 00 00 is required for the amended claims presented and has been calculated as shown on page 3.
Please charge Deposit Account No. 23-0450 in the amount of the additional fee above, or such greater or lesser amount of excess fees for claims as the Commissioner determines is required by law. This letter is submitted in triplicate.
PO11182 08/16/90 07384187 23-0450 110 114 180.00CH
PLD-115 Page 1 of 3 Pages 4/89 PD-No: 3844-01-JT
Applicant(s) hereby petitions for a two(2) month extension of the period for response in the above-captioned application which expired on June 22 1990 and hereby authorizes the Commissioner to charge the extension fee of $180.00 , or such greater or lesser amount as may be required by 37 CFR 1.17, to Deposit Account No. 23-0450. This transmittal letter is submitted in triplicate. If for any reason the extension requested above is insufficient to extend the period for response to the date of this paper, applicant(a) hereby petitions far an extension of time under 37 CFR 1.136(a) sufficient to obtain such an extension and/or the revival of the above-captioned application as having been unintentionally abandoned and authorizes the Commissioner to charge the required fees under 37 CFR 1.17 to Deposit Account No. 23-0450.
Respectfully submitted,
Date
2800 Plymouth Road Ann Arbor, MI 48105 Tel. (313)996- 7190
Attachments: Reply/Amendment Form PLD-115 (in triplicate) Calculation of Additional Fees for Amended Claims
PLD- 115 Page 2 of 3 Pages 4/89 PD -NO :3844-01-JT
CALCULATION OF ADDITIONAL FEES FOR AMENDED CLAIMS
Applicant(s) have calculated the additional fees for the amended claims as follows :
Total Claims 00.00
claims 1 minus 00.00
* If entry in Column 2 is less than entry in Column 4, enter "0" in Column 5.
If claims previously paid for are less than 20 for total claims or 3 for independent claims, enter 20 for total claims or 3 for independent claims in Column 4.
PLD- 115 Page 3 of 3 Pages 4/89 ,
IN THE united STATES patent AND TRADEMARK OFFICE
APPLICANT BRUCE DAVID ROTH EXAMINER: R. HOWARD SERIAL NO.: 07/384,187 GROUP ART UNIT: 121
JULY 21, 1989 PAPER NO.: 4
FOR
ACID, ITS LACTONE FORM AND SALTS THEREOF
August 6,1990 i ("3
amendment AND remarks
Hon. Commissioner of Patents and Trademarks Washington, D-C. 20231
Dear Sir:
This is response to an Office Action dated March 22, 1990 in the above noted application.
Therefore, please enter the following amendments:
- acid or (2R-trans)-5- 07/384,187 - PD 8 44- 1 I A compound of claim which is R- (R*R*) 1-2- 'II -3-phenyl-4- (phenyl , acid.
-2- 4-diphenyl
Claims 1-12 are rejected under 35 USC 1.12, second paragraph. Applicant gratefully acknowledges the specificity of Examiner's comments regarding this rejection. Applicants
Claims 1-12 are also rejected under 35 USC 102(b) as being anticipated by Roth, U.S. Patent. 4,681,893 (hereinafter Roth) .
contrary to Examiner's reading roth does not teach the presently claimed substituted -4- y an- 2 - e the corresponding dihydroxy acids .
The presently claimed compounds are for individual enantiomers and. therefore differ from the teaching in Roth only I to to mixtures of enantiomers. I This is evident from the stereochemistry of Roth's
' disclosure at: column 3 lines 49 to 54 and column 6 lines 56-58 note this regard i which contrary to Examiner's in only indicates I a trans-isomer and an R*R* configuration
I
I I 07/384,187 -3- PD-3844-01-JT
i Roth the nomenclature cited by Examiner; i .e., "trans-isomers" and "R*R* configuration", expresses a relationship between two substituents on adjacent assymetric carbons recognized as one of two possible isomers, i.e., compounds having the two substituents either on the same side of adjacent assymetric carbons or on the opposite sides of adjacent assymetric carbons. Thus in Roth "trans" indicates an isomer having the relationship as an opposite sides and "R*R*" indicates an isomer having the relationship as on the same side.
However, additionally in molecules of the kind disclosed in Roth each possible isomer also exists in two forms which depend on a configuration which is expressed in absolute terms relative to the remainder of. the molecule. The forms are denoted as an R form and an S form. These two forms are recognized by an ordinarily skilled artisan to be enantiomeric forms each having a specific chirality. In Roth the disclosure is not limited to compounds having such a specific chirality. Thus, each isomer of Roth is a mixture of enantiomers and not the currently claimed individual enantiomers having an R chirality.
Specifically, Roth' s difference from the present claims is readily recognized by the term "R-transi somer" and the term "R( R*R* ) in the instant claims compared to the Ianguage Examiner found in Roth which is also noted above. This difference indicates the present claims are for individual enantiomers having a pair of substituents in each case with an absolute relationship to the molecule not previously disclosed, and, further, each member of the pair has a relationship to each other
In other words, roth does disclose the relationship between each member of a pair of substituents but is silent regarding chirality within the enantiomer or ab so 1. ute relationship of the pair to the molecule leaving the Roth disclosure limited to a mixture of enantiomers.
For purposes of clarifying the above discussion it is noted. here that the onantiomer not now in the claims would be 07/384,187 -4- PD-3 8 44 - 0 1 JT recognized as an "S-trans-isomer" and an "S(R*S*)" enantiomer respectively.
Clearly the above discussion obviates the rejection of record.
For the above reasons reconsideration and allowance is requested for claims 4-15 which are all the claims now in the present application.
Respectfully submitted, Bruce David Roth By his Attorney
Assistant Patent Counsel (Reg. No. 29,450) Warner-Lambert Company Pharmaceutical Research Division 2800 Plymouth Road Ann Arbor, Michigan 48105 (313) 996-7190
JT1S3502.DOC UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office Address : COMMISSIONER OF PATENTS AND TRADEMARKS Washington. D.C. 20231
E 384401JT
HALEY J
121
1 1 13/90
This application has been examined Responsiveto communication filed on action Is made final.
A shortened statutory period for response to this action Is set to expire three month(s). from the date of this letter. Failure to respond within the period for responsewill cause the application to become ebandoned. 35 U.S.C. 133
Part I THE FOLLOWING attachment(s) ARE PART OF THIS ACTION
1. Notice of References Cited by Examiner, PTO-892. 2. Notice re Patent Drawing, PTO-948. 3. Notice of Art Cited by Applicant, PTO-1449. 4. Notice of informal Patent Application. Form PTO-152. 5. information on HOW to effect Drawing Changes, PTO-1474. 6.
Part SUMMARY OF ACTION
claims 4 15 are pending in the application.
Of the above, claims are withdrawn from consideration.
2 claims 1-3 have been cancelled.
claims are allowed.
4. are rejected.
5. Claims are objected to.
6. Claims are subject to restriction or election requirement.
7. This application has been filed with informal drawings under 37 C.F.R. 1.85 whlch are acceptable for examination purposes.
8. Formal drawings are required in response to this Office action,
9. The corrected or substitute drawings have been received on . Under 37 C.F.R. 1.84 these drawings are acceptable. not acceptable (see explanation or Notice re Patent Drawing, PTO-948).
10. The proposed additional or substitute sheet(s) of drawings, filed on has (have) been approved by the examiner. disapproved by the examiner (see explanation).
11. The proposed drawing correction, filed on has been approved. disapproved (see explanation).
12. Acknowledgment is made of the claim for priority under U.S.C. 119. The certified copy has been received not been received
been filed in parent application, serial no. filed on
13. Since this application appears to be condition for allowance except for formal matters, prosecution as to the merits Is closed In accordance with the practice under Ex parte Quayle, 1935 C.D. 11: 453 O.G. 213.
14. Other
EXAMINER'S ACTION PTOL-326 (Rev. 6-88) . .
Serial No. 07/384,187 -2-
Art. unit 121
Claims 4-15 are pending. The following is a quotation of the appropriate paragraphs of 35 U.S.C § 102 that form the basis for the rejections under this section made in this office action:
A person shall be entitled to a patent, unless -- (In) the invention was patented or described in a printed publication in this or a foreign country or in public use or on sale in this country more than one year prior to the date of application for patent in the United States.
Claims 4-15 are rejected under 35 U.S.C. § 102(b) as being anticipated by Roth, us Patent: 4,681,893. Roth teaches the claimed substituted alkyl] 4-
which are sufficiently limited in number, the reference is deemed .
Serial No. 07/384,187 -3- art. unit 123
Claim 13 is rejected under 35 U.S.C. § 112, second
paragraph, as being indefinite for failing to particularly point
regards as the invention. The claim should be drawn to a single invention. The term ‘’or‘’should be substituted for “and” to claim a single invention. Applicant’s amendment necessitated the new grounds of rejection Accordingly, THIS ACTION is MADE FINAL. See M.P.E.P.
§ 706.07(a). Applicant: is reminded of the extension of time policy as set forth in 37 C.F.R. § 3.336(a).
A SHORTENED STATUTORY PERIOD FOR RESPONSE TO THIS FINAL ACTION IS SET TO EXPIRE, THREE MONTHS FROM THE DATE OF THIS ACTION. IN THE EVENT A FIRST RESPONSE is filed WITHIN TWO MONTHS OF THE mailing date OF THIS FINAL ACTION and THE ADVISORY ACTION us NOT mailed AFTER THE END OF THE THREE-MONTH SHORTENED STATUTORY PERIOD, THEN THE SHORTENED STATUTORY PERIOD WILL EXPIRE ON THE DATE THE ADVISORY ACTION is MAILED, AND ANY EXTENSION FEE PURSUANT TO 37 C.F.R. § 1.136 (a) WILL BE calculated FROM THE mailing DATE OF THE advisory ACTTON. IN NO EVENT WILL THE STATUTORY PERIOD”) FOR RESPONSE expire LATER THAN SXX MONTHS FROM THE DATE OF THIS final ACTTON. CLAIMS AS FILED PART I other THAN A small ENTITY SMALL ENTITY
NO FILED EXTRA
TOTAL
CLAIMS AS AMENDED PART
OTHER THAN A SMALL entity SMALL entity
minus .. I I I
TOTAL
PRESENT OR EXTRA
120 I
OR TOTAL
TOTAL TOTAL addit FEE
I i mi
0
0 C 2
0
1 IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
Art Unit 121 Examiner J. Haley applicant(s) Bruce David Roth Serial Number : 07/384,187 Filed july 21, 1989 For ( CARBONYL] FORM AND SALTS THEREOF Commissioner a f Paten t s trademarks Washington, D.C. 20231 ATTENTION: BOX FWC REQUEST FOR FILE WRAPPER CONTINUING APPLICATION UNDER 37 CFR 1.62
Sir:
Applicant(s) hereby request that a continuation divisional continuation-in-part application be filed in connection with the abandonment of the above-captioned application in accordance with the provisions of 37 CFR 1.62. The particulars of this request are set forth below:
This paper is being filed under the provisions of 37 CFR 1.10 and contains hereon the required Certificate of Mailing by "Express Hail".
RETROACTIVE EXTENSION OF TIME (37 CFR 1.136(a)) OR REVIVAL (37 CFR 1.137(b)) IN PARENT APPLICATION
Applicant(s) hereby petition for a ONE month extension of the period for response in the above-captioned application which expired on FEBRUARY 13 1991 and hereby authorize the Commissioner to charge the extension fee of $100.00 , or such greater or lesser amount as may be required by 37 CFR 1.17, to Deposit Account No. 23-0450. If for any reason the extension requested above is insufficient to provide co-pendency under 35 USC 120, Applicant(s) hereby petition for an extension of time under 37 CFR 1.136(a) sufficient to obtain co-pendency and/or the revival of the above-captioned application as having been unintentionally abandoned and authorize the Commissioner to charge the required fees under 37 CFR 1.17 to Deposit Account No. 23-0450. PLD 6 2 Page 1 of 3 11/90 AMENDMENTS IN CONNECTION WITH FILING
A. [ Please enter the unentered amendment(s) made after final rejection (37 CFR 1.116) in the above-captioned application dated
B. Please enter the preliminary amendment provided herewith.
C. [ Please cancel claims
III. FILING FEES
A. The filing fee has been calculated as shown below:
... CLAIMS PENDING AFTER AMENDMENTS
B. SPECIFIC deposit account authorization Please charge my Deposit Account No. 23-0450 in the amount of the total filing fee above. Triplicate copies of this paper are enclosed.
C. GENERAL deposit ACCOUNT authorization The Commissioner is hereby authorized to charge payment of the following fees associated with this communication or during the pendency of this application or credit any overpayment to Deposit account No. 23-0450.
(1) Any additional filing fees or fees for the presentation of additional claims required under 37 CFR 1.16.
(2) Any patent application processing fees under 37 CFR 1.17.
No authorization is given to charge the Issue Fee (37 CFR 1.18).
D. [ A check in the amount of $ to cover the filing fee is enclosed.
PLD - 62 Page 2 of 3 11/90 PATENT/Docket No: 3844-C1-0l-JT
IV. ADDITIONAL INFORMATION
A. A new declaration or oath is attached pursuant to 37 CFR 1.62(c).
The new application is a continuationor divisional application which discloses and claims only subject matter disclosed in the above- captioned application and no new oath is required, 37 CFR 1.62(c).
B. The title of the above-captioned application as originally filed, if different from the last amended title set forth above, is as follows:
C. [ The name of the Applicant(s) as originally filed, if different from the last amended Applicant(s) set forth above, is as follows:
D. [ The current correspondence address of the Applicant(s), if different from that set forth in Applicant(s) declaration (37 CFR 1.63) is as f 01lows :
E. [ Priority under 35 USC 119 is claimed For the foreign applications identified as follows:
Respectfully submitted,. '
+-- I Date: FEBRUARY 1991 . , Registration No, 29 450 Warner-Lambert company ' , 2800 Plymouth Road Ann Arbor, HI 48105 Tel. No. (313) 996-7190
Attachments: Form PLD-62 (in triplicate) New Declaration or Oath Preliminary Amendment
PLD-62 Page 3 of 3 11/90 United united STATES patent AND TRADEMARK OFFICE
APPLICANT BRUCE DAVID ROTH EXAMINER:
SERIAL NO.: GROUP ART UNIT: FILED: PAPER NO. :
FOR : [R-(R*R*)]-2-
ACID, ITS LACTONE FORM AND SALTS THEREOF
! February 26, 1991 !
PRELIMINARY AMENDMENT AND REMARKS
Hon. Commissioner of Patents and Trademarks washington D.C. 20231.
Dear Sir:
This is a Preliminary Amendment in the above-noted application which is a File Wrapper Continuing Application Under 37 C.F.R. 1.62 of United States Serial No. 07/384,187 filed July 21, 1989, therefore, please enter the following amendment :
In Claim 13, line 5 change "and" to --or--.
i On page 1, on the line after "AND SALTS THEREOF" add . . ---This in a continuation ofU.S. application Serial number 07/384,187 filed july 21, 1989, nowpending - 2 .-
Claims 4-15 are rejected under 35 U.S.C. 102(b) as being anticipated by Roth. Examiner cites 197 USPQ 5 (CCPA 1978) to support this rejection alleging that "(w)here a reference discloses a genus of compounds of similar structure which are sufficiently limited in number, the reference is deemed to provide description of those compounds just as specifically as if they were identified by name. Examiner further states that "one merely has to select from the limited possibility of isomers to arrive at the claimed invention, and separate them using conventional techniques" thereby apparently admitting that Roth indeed discloses an isomer which is in turn a mixture of enantiomers. On this basis Shaumann is not the applicable case law because in shaumann, although a formula in the cited art was argued, the issue was not one relating to a mixture of enantiomers but to whether the cited art provides sufficient enabling disclosure to support the formula. However, in the same volume of the USPQ applicant respectfully requests that Examiner's attention be directed to In May and eddy 197 USPQ 601 at 607 as follows:
As recognized in In re Williams, 36 CCPA 756, 171 F.2d 319, 80 USPQ 150 (1948)) the novelty of an optical isomer is not negated by the prior art disclosure of its racemate.
Williams applied this statement to find isomers novel over a cited disclosure to racemates. Clearly this case law is applicable e here
Following the case Applicant also how provides by a declaration a comparison among each enantiomer and mixture of enantiomers. This comparison is provided to overcome the Roth reference of the present rejection to facilitate a finding of patentability and moving the prosecution toward resolution of pertinent issues. in other words, although examinar has not included a rejection under 35 U.S.C. 103 Applicants are Including a rebuttal- of such a rejection to comply with the
...Williams , , . . .. , case law. -3-
The declaration submitted with the present Remarks provides the data as net out in the present application in a manner to provide patentability to the application. This data shows a difference in kind not degree between the activity of each enantiomer and between the activity of an enantiomer and the mixture of enantiomer. In other words, the declaration is submitted to provide evidence of patentability to the instant invention.
Claim 13 is rejected under 35 U.S.C. 112, second paragraph, in which Examiner suggested changing "and" to --or-- to claim a single. Applicant's compliance with this suggestion obviates this rejection.
For the above reasons reconsideration and allowance is requested for Claims 4-15 which are now all the claims in the application.
Respectfully submitted, Bruce David Roth By his Attorney'
Assistant Patent Counsel (Reg. No. 29,450) Warner-Lambert Company Pharmaceutical Research Division 2800 Plymouth Road Ann Arbor, Michigan 48105 (313) 996-7190
JT1S3683 .DOC
ATTACHMENT: DECLARATION UNDER 37 CFR 1.132 ( bruce DAVID roth PD-3844-C1-01-JT
APPENDIX A
APPENDIX A IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
APPLICANT BRUCE DAV ID roth EXAMINER :
SERIAL NO.: GROUP ART UNIT:
FILED: PAPER NO. :
FOR :
ACID, ITS LACTONE FORM AND SALTS THEREOF
February 25, 1991
DECLARATION under 37 CFR 1.132 hon Commissioner of Patents arid Trademarks Washington, D.C. 20231
Dear Sir:
I, Bruce D. Roth, declare as follows:
THAT, I am the inventor in the above identified application.
2. THAT, I have been awarded the degrees of Bachelor of Science in Chemistry by the St. Joseph's College, Philadelphia, PA in 1976, and a Doctor of Philosophy in Organic Chemistry by Iowa State University at Ames, IA in February 1981;
3. THAT, I was a Postdoctoral Fellow under the direction of Professor Andrew S. Kende at the University of Rochester, Rochester, NY from December 1981. to April 1982;
4. THAT, I am currently Section Director, Chemistry Department, Parke-Davis Pharmaceutical Research Division, Warner-Lambert Co. after receiving increasingly responsible positions from my employment in 1982; -2- PD-3844-C1-0l-JT
5. THAT, I am a member of the American Chemical Society and have presented numerous papers and have numerous publications and patents a5 shown in Appendix A attached;
6. THAT, I and persons working under my direction engage in a research program on the treatment of humans suffering from hypercholesterolemia ;
7. THAT, as part of said research program the antihyper- cholesterolemia properties of
I
I
pyrrole-1-heptanoic acid hemicalcium salt (I)
pyrrole-1-heptanoic acid hemicalcium salt (II) and mixtures thereof are assessed using essentially the CSI screen that is disclosed in U.S. patent No. 4,681,893, attached as Appendix B;
8. THAT, in said assessment, the datum from the Compound I, the datum from its enantiomer the Compound II and the datum from the racemate of the two compounds I and II are as follows:
I R- (R*R* ) isomer 0.025 II [S-(R*R*)] isomer >1.00 Racemate 0.26
9. THAT, the data demonstrate that the Compound I provides an IC which indicates activity greater than 50 fifty-fold more than that of Compound II and which indicates activity at least ten--fold more than that of the racemate; -3- PD-3844-C1-01-JT
10. THAT, the differences in the data of paragraph 9 among Compounds I, II and racemate shows the activity of Compound I is surprising and unexpected because if the Compound II is accepted as inactive, the activity of the Compound I would be expected to be only accepted as twice that of the racemic mixture;
11. THAT, all statements made herein of my own knowledge are true and that all statements made on information are believed to be true, and further that these statements are made with the knowledge that willful false statements and the like so made are punishable by fine or imprisonment, or both, under Section 1001, Title 18, of the United States Code and that such willful- false statements may jeopardize the validity of the above identified US patent application having Bruce David Roth as inventors for
HEPTANOIC ACID, ITS LACTONE FORM AND SALTS THEREOF or any patent issuing thereon.
Bruce D. Roth
JT1S3684.DOC PD-3844-C1-01-JT
APPENDIX B
APPENDIX B PROFESSIONAL SOCIETIES
American Chemical Society
PAPERS PRESENTED
1. B. Roth, M. Hoefle, D. Sliskovic, C. Stratton, M. Wilson, and R. Newton. "Trans-6- a new class of HMG-CoA reductase inhibitors" 194th National ACS Meeting, New Orleans, LA August 24-29, 1987.
2. B, Roth. "The Pharmacological Properties and Structure-ActivityRelationships of a New Series of ACAT Inhibitors" 1989 Gordon Conference on Medicinal Chemistry, New London, NH August 2, 1989.
3. W. H. Roark, B. K. Trivedi, A. Holmes, B D. Roth, C. J. Blankley, M. L. Hoefle, B R, Krause K. Kieft and R. L. Stanfield. "SAR Studies on Novel Fatty Acid Anilide ACAT Inhibitors: Potential Hypocholesterolemic Agents" 1 98th ACS National Meeting, Miami Beach, FL September 10-15, 1989.
4. W. H. Roark, B. D. Roth, A. Holmes, K. Kieft, A. D. Essenburg, R. L. Stanfield and B R. Krause. "Modifications of Oleic Acid Anilide ACAT Inhibitors: Replacement of the Amide Bond 198th ACS National Meeting, Miami Beach, FL September 10-15, 1989.
5. J. A. Picard, D R. Sliskovic, B. D Roth and R. S. Newton. "Trans-6-[2- A New Class of HMG-CoA Reductase Inhibitors" 198th ACS National Meeting, Miami Beach, FL September 10-15, 1989.
6. B. R. Krause, M. L Hoefle, A. Holmes, 8. D. Roth, K. A. Kieft. P. D Uhlendorf, R. L. Stanfield, C. S. Sekerke, T, M. A. Bocan and R. S. Newton. "Lipid- Regulating Activity of PD 128042, A Novel Inhibitor of Acyl-CoA: Cholesterol (ACAT)" X International Symposium on Drugs Affecting Lipid Metabolism, Houston, TX November 8-1 1, 1989.
7. T. M. A Bocan, S. BakMueller, P. D. Uhlendorf, E. Ferguson, A. Holmes, B. D Roth and R. S. Newton, "Effect of ACAT Inhibitors and Selected Lipid Lowering Agents on Atherosclerotic Lesions in a Cholesterol-Fed Rabbit Model" X International Symposium on Drugs Affecting Lipid Metabolism. Houston, TX November 8-1 1, 1989. 2 8. R. s. Newton, B. R. Krause, R. L. Stanfield, T. M. A. Bocan, M. K. Shaw, E. Ferguson, C. Sekerke, B D Roth and D. R. Sliskovic. "Pharmacological Profile of PD 134298-38A, A Novel HMG-CoA Reductase Inhibitor" X International Symposium on Drugs Affecting Lipid Metabolism. Houston, TX November 8-1 1, 1989.
9. M. W. Creswell, A. W. Chucholowski, B. D. Roth, C. J. Blankley, E. Ferguson, M. K. Shaw, B. R. Krause and R. S. Newton. and and and their N-oxides, A New Class of HMG-CoA Reductase Inhibitors." 22nd ACS Regional Meeting, Saginaw, MI June 6-8, 1990.
10. J. A. Picard, E. Ferguson, B. R. Krause, R. S. Newton, W. H. Roark, B. D. Roth, M. K. Shaw and 0. R. Sliskovic. N-Heteroaryl Pyrazole Mevalonolactones, Potent Inhibitors of HMG-CoA Reductase Both In Vitro and In Vivo" 200th ACS National Meeting, Washington D.C. August 26-31, 1990.
11. D. R. Sliskovic, C. J. Blankley, T. M. A. Bocan, A. W. Chucholowski, P. L Creger, M. W. Creswell, E. Ferguson, P. O'Brien, J. A Picard, W. H. Roark, B. D. Roth and M. W. Wilson. Tissue Seletivity of HMG-CoA Reductase Inhibitors" 200th ACS National Meeting, Washington D.C. August 26-31, 1990. 12. W. H. Roark, A, Holmes, B. D. Roth, B. K. Trivedi, A. D. Essenburg, K. Kieft, B. R. Krause and R. L. Stanfieid. Inhibitors of Acyl-CoA Cholesterol Acyltransferase (ACAT" 200th ACS National Meeting, Washington D.C. August 26-31, 1990.
PAPERS
1. George A. Kraus and Bruce Roth. "Michael Addition Reaction of Angelica Lactone." Tetrahedron Letters 31 29 (1977).
2. George A. Kraus and Bruce Roth. "Rearrangements of Acyloxyfurans and Thiophenes." Journal of organic Chemistry, 43, 2072 (1978).
3. George A. Kraus and Bruce Roth. "Synthesis of Pyrano-&Lactone Antibiotics. 1. Synthesis of Journal of Organic Chemistry, 43. 4923 (1 978).
4. George A. Kraus and Bruce Roth. "Synthetic Studies Toward Verrucarol. 2. Synthesis of the AB Ring System.' Journal of Organic chemistry 45. 4825 (1980). 5. George A. Kraus, Kevln A Frazier, Bruce D. Roth, Michael J. Taschner, and Kent Neuenschwander. "Conversation of Lactones into Ethers" Journal of Organic Chemistry,46, 241 7 (1981). 3 6. George A. Kraus, Hidetsura Cho, Steven Crowley, Bruce Roth, Hirohiko Sugimoto. and Susan Prugh. "Phthalide Annulation: The Synthesis of Kalafungin, Pachybasin, and Chrysophanol." Journal of Organic Chemistry. 48, 3439 (1 983).
7. George A. Kraus, Bruce Roth, Kevin Frazier, and Masayuki Shimagaki. "Stereoselective Synthesis of Calonectrin." Journal of the American Chemical Society, 104, 11 14-1116 (1982).
8. Andrew S. Kende and Bruce Roth. "StereoselectiveTotal Synthesis of Ajugarin IV." Tetrahedron Letters 1751-1 754 (1982).
9. Andrew S. Kende, bruce Roth, and Pauline J. Sanfilippo. "Facile. Palladium (II) Mediated Synthesis of Bridged and Spirocyclic Bicycloalkenones." Journal of the American Chemical Society, 104, 1784-1785 (1982). 10. Andrew s. Kende, Bruce Roth, Pauline J. Sanfilippo, and Thomas J. Blacklock "Mechanism and Regioisomeric Control in Palladium (II) Mediated Cycloalkenylations. A Novel Total Synthesis of (±)-Quadrone." Journal of the American Chemical Society, 1045808-5810 (1982). 11. Bruce D. Roth and W. Howard Roark. "Synthesis of a Chiral Synthon for the Lactone Portion of Compactin and Mevinolin." Tetrahedron Letters, 29- 1255-1258 (1988). 12. Bruce D. Roth, Daniel F. Ortwine, Milton L Hoefle, Charlotte D. Stratton, Drago R. Sliskovic, Michael W. Wilson, and Roger S, Newton. "Inhibitors of Cholesterol Biosynthesis. Part I. A Novel Series of HMG-CoA Reductase Inhibitors. 1. Effects of Structural Modifications at the Two and Five Positions of the Pyrrole Nucleus. J. Med. Chem 33.21-31 (1990).
13. Drago R. Sliskovic, Bruce D. Roth, Michael W. Wilson, Milton L Hoefle, and Roger S. Newton. "Inhibitors of Cholesterol Biosynthesis. Part 2. 1,3,5-Trisubstituted- J. Med. Chem., 33, 31 -38 (1990).
14. Mary Kay Shaw, Roger S. Newton, Drago R. Sliskovic, Bruce D. Roth, Erika Ferguson, and Brian R. Krause. "Hep-G2 Cells and Primary Rat Hepatocytes Differ in their Response to Inhibitors of HMG-CoA Reductase." Biochem. Biophys. Res. Comm., 170, 726-734 (11990
15. bruce D. Roth, C. John Blankley, Alexander W. Chucholowski, erika Ferguson, Milton L Hoefle, Daniel F. Ortwine, Roger S. Newton, Cathy S. Sekerke, Drago R. Sliskovic, Charlotte D. Stratton, and Michael W. Wilson. "Inhibitors of Cholesterol Biosynthesis. Part 3. Inhibitors of HMG-CoA Reductase. 2 Effects of introducing Substituents at Positions Three and Four of the Pyrrole Nucleus." J. Med. Chem., 34 0000 (1991). 4 16. Drago R. Sliskovic, Joseph A. Picard, W. Howard Roark, Bruce D. Roth, Erika Ferguson, Brian R. Krause, Roger S. Newton, Cathy Sekerke, and Mary Kay Shaw. "Inhibitors of Cholesterol Biosynthesis. Part 4. A Novel Series of HMG-CoA Reductase Inhibitors." J. Med. Chem. 34, 0000 (1991).
17. Bruce D. Roth, Thomas M. A. Bocan, C. John Blankley, Alexander W. Chucholowski, Paul L. Creger, Mark W. Creswell, Erika Ferguson, Roger S. Newton, Patrick O'Brien, Joseph A. Picard, W. Howard Roark, Catherine S. Sekerke, D. Robert Sliskovic, and Michael W. Wilson. "The Relationship Between Tissue Selectivity and Lipophilicity for Inhibitors of HMG-CoA Reductase." J. Med. Chem. 34, 0000 (1991).
PATENTS
1. Inhibitors of Cholesterol Synthesis. Bruce D. Roth, Milton Hoefle, and Charlotte Stratton. USP 4,647,576.
2. or Inhibitors of Cholesterol Synthesis. Bruce D. Roth. USP 4,681,893.
3. -4.) -1 Inhibitors of Cholesterol Biosynthesis. Bruce D Roth and Drago R. Sliskovic. USP 4,735,958. 4. Saturated Fatty Acid Amides as Inhibitors of Acyl-CoA: Cholesterol Acyltransferase. Milton L Hoefle, Ann Holmes, and Bruce D. Roth. USP 4,716,175.
5. Saturated fatty Acid Amides as Inhibitors of Acyl-CoA Cholesterol Acyltransferase. Milton L Hoefle, Ann Holmes, and Bruce D. Roth. USP 4,743,605.
6. Treating Fungal Infections with Carl L Heifetz, Milton L Hoefle, Bruce D. Roth, Drago R. Sliskovic, and Michael W. Wilson. USP 4,751,229.
7. or hydroxypyran-2-one Inhibitors of Cholesterol Bruce D. Roth and Drago R. Sliskovic. USP 4,808,621. 8. Inhibitors of Cholesterol Biosynthesis. Alexander W. Bruce D. Roth and Drago R. Sliskovic. USP 4,868,185.
9, 6-(( and Inhibitors of Cholesterol Biosynthesis. Alexander W. Mark W. Creswell, Bruce D. Roth and Drago R. Sliskovic. USP 4,906,624. 5 1 0. Bicycloheptane and Bicyclooctane Substituted Inhibitors of Cholesterol Biosynthesis. Bruce D. Roth. USP 4,957,940.
1 1 . Trans-6-( 2-( hydroxypyran-2-one Inhibitors of Cholesterol Biosynthesis. Joseph A. Picard, Bruce D. Roth and Drago R. Sliskovic. USP 4,957,971.
REVIEW ARTICLES
1. bruce D Roth, Drago R. Sliskovic and Bharat K. Trivedi. "Treatment of Hypercholesterolemia." Annual Reports in Medicinal Chemistry 24,147-1 56 (1 989).
P :\C .CV .IN THE UNITED STATES patent AND trademark OFFICE
Examiner : applicant(s) BRUCE DAVID ROTH Serial Number : Filed : POP [R- (R*R*) -2- METHYLETHYL-3-PHENYL-4- (PHENYLAMINO)CARBONYL] IH-PYRROLE-1-HEPTANOIC ACID, ITS LACTONE FORM commissioner of Patents and Trademarks AND SALTS THEREOF Washington, DC 20231
reliance on PRIOR information disclosures statement under 37 CFR 1.97 AND 1.98 IN THE PRESENT FILE WRAPPER
Sir : For the lnformation disclosure statement under 37 CFR 1.97 and 1.98 In the present application, applicant relies on the prior information disclosure Statement(s) filed on JULY 21, 1989 in Serial No. 07/384,187 , filed JULY 21, 1989 , which is contained in the file wrapper of the present application pursuant to 37 CFR 1.62. Respectfully submitted ,
FEBRUARY 26, 1991 JOAN E attorney Date ,/' Reg- registration NO. 29,450 Telephone lumber: (313) 996-7190
PLD-97.3 1012511 985 PATENT/Doc No 3844-01-JT
N THE UNITED STATES PATENT AND TRADEMARK OFFICE
t
Serial Number :
Far : [R-R*R*) -2- (4-FLUOROPHENYL) ENYLAMINO) ACID, ITS LACTONE FORM AND SALTS THEREOF Commissioner of Patents and Trademarks Washington, D.C. 20231 INFORMATION DISCLOSURE STATEMENT UNDER 37 CFR 1.97 AND 1.98 Sir: Provided herewith are copies of patents and publica- tions as listed in the attached form PTO-1449. A concise explanation of the relevance of each reference listed in Form PT0-1449 is: [XI contained in the above-captioned specification as filed; and/or provided in a communication enclosed with this paper. Applicant respectfully asserts that the substantive provisions of 37 CFR 1.97 and 37 CFR 1.98 are met by the provision of the foregoing statement. Respectfully submitted,
JULY 21, 1989 ELIZABETH M. ANDERSON , AGENT Date Registration N0.31,585 warner-lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 Tel. NO. (313)996-7304
Attachment: PTO-1449
PLD-97.1 11/87 I I I 1
translation documentnumber DATE country subclass yes no
I UNITED state DEPARTMENT OF COMMERCE Patent and trademark Office Address COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231
... . .
This application has been examined to communication filed on This action le made final.
A shortened statutory period for response to this action is set to expire month(s), days from the date of this letter. Failure to respond within the period for responsewill cause the application to becomeabandoned. 35 U.S.C. 133 Part I THE FOLLOWING attachment(s) ARE PART OF THIS ACTION:
1, Notice of References Cited by Examiner, PT0-892. 2. Notice re Patent Drawing. PTO-948. 3. Notice of Art Cited by Appliant, PTO-1449. 4. Notice of Informal Patent Application. Form PTO-152 5. Information on How to Effect Drawing Changes, PTO-1474. 6.
Part SUMMARY OF ACTION
1. Claims 4 are pending in the application.
Of the above, claims are withdrawn from consideration.
Claims '-3 haw been cancelled.
3.0 Claims are allowed.
are rejected.
5. Claims are objected to.
6. Claims are subject to restriction or election requirement.
7. This application has been filed with informal drawings under 37 C.F.R. 1.85 which are acceptable for examination purposes.
8. Formal drawings are required in response to this Office action.
The corrected or substitutedrawings have been received on . under 37 C.F.R. 1.84 these drawings are acceptable; not acceptable (see explanation or Notice re Patent Drawing, PTO-948).
10. The proposed additional or substitute sheet(s) of drawings, filed on . has (have) been approved by the examiner; disapproved by the examiner (see explanation).
The proposed drawing correction, filed , has been approved; disapproved (see explanation).
12. Acknowledgement is made of the claim for priority under U.S.C. 119. The certified copy has been received not been received been filed in parent application,serial no. filed on
Since this application appears to be in condition for allowance except for formal matters prosecutionas to the merits Is closed in accordance with the practice under Ex parte Quayle, 1935 C.D. 11453 O.G. 213.
14. Other
EXAMINER'S ACTION PTOL-326 (Rev.9.89) Serial No. 07/660976 -2-
Art Unit 121
Claims 4-15 are pending. The prior art made of record and not relied upon is considered pertinent to applicant's disclosure. Applicant's Information Disclosure Statement is noted. However, the references cited therein are already of record. The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action: A person shall be entitled to a patent unless -- (b) the invention was patented or described in a printed publication in this or a foreign country or in public use or on sale in this country, more than one year prior to the date of application for patent in the United States.
Claims 4-15 are rejected under 35 U.S.C. § 102(b) as being anticipated by Roth, U.S. Patent 4,681,893 for all of the reasons of record in Paper No. 3 and 6. Applicant's arguments have been very carefully considered, however such are not persuasive. Applicant urges that In re Schaumann is not applicable case law. However, Schaumann is seen to apply since such a limited number of possible forms of the isomer exists that a sufficient description of each isomer is provided by the reference. This is especially considered so due to the overt statement of contemplated isomers appearing at column 4, lines 45-54. Therefore, the rejection under 35 USC
102(b) under 35 USC 103, the Roth declaration is not pertinent. Serial No. 07/660976 -3- Art Unit 121
This is a continuation of applicant's earlier application
S.N. 07/384,187. All claims are drawn to the same invention claimed in the earlier application and could have been finally rejected on the grounds or art of record in the next Office action if they had been entered in the earlier application.
Accordingly, THIS ACTION IS MADE FINAL even though it is a first action in this case. See M.P.E.P. § 706.07(b). Applicant is reminded of the extension of time policy as set forth in 37
C.F.R. § 1.136(a).
A SHORTENED STATUTORY PERIOD FOR RESPONSE TO THIS FINAL ACTION IS SET TO EXPIRE THREE MONTHS FROM THE DATE OF THIS ACTION. IN THE EVENT A FIRST RESPONSE IS FILED WITHIN TWO MONTHS OF THE MAILING DATE OF THIS FINAL ACTION AND THE ADVISORY ACTION IS NOT MAILED UNTIL AFTER THE END OF THE THREE-MONTH SHORTENED STATUTORY PERIOD, THEN THE SHORTENED STATUTORY PERIOD WILL EXPIRE ON THE DATE THE ADVISORY ACTION IS MAILED, AND ANY EXTENSION FEE PURSUANT TO 37 C.F.R. § 1.136(a) WILL BE CALCULATED FROM THE MAILING DATE OF THE ADVISORY ACTION. IN NO EVENT WILL THE STATUTORY PERIOD FOR RESPONSE EXPIRE LATER THAN SIX MONTHS FROM THE DATE OF THIS FINAL ACTION.
HALEY :m' September 16, 1991 UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office Address: COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231 both
1
' EXAMINER INTERVIEW SUMMARY RECORD
All participants (applicant, applicant's representative. PTO personnel):
Date of interview ' .
Type Telephonic Personal (copy is given to applicant applicant's representative).
Exhibit shown or demonstration conducted: Yes No. If yes. brief description:
......
Claims discussed ......
Identification of prior art discussed: ......
......
(A fuller description, if necessary, and a COPY of the amendments. if available, which the examiner agreed would render the claims allowable must be attached. Also, where no copy of the amendments which would render the claims allowable is available, a summary thereof must be attached.)
Unless the paragraphs below have been checked to indicate to the contrary, A FORMAL WRITTEN RESPONSE TO THE LAST OFFICE ACTION IS NOT WAIVED AND MUST INCLUDE THE SUBSTANCE OF THE INTERVIEW (e.g., items 1-7 on the reverse side of this form). If a response to the last Office action has already been filed, then applicant is given one month from this interview date toprovide a statement of the substance of the interview.
It is not necessary for applicant to provide a separate record of the substance of the interview.
Since the examiner's interview summary above (including any attachments) reflects a complete response to each of the objections, rejections and requirements that may be present in the last Office action, and since the claims are now allowable, this completed form is considered to fulfill the response requirements of the last Office action.
PTOL-413 (REV 1.84) examinar’s PATEN ocket No:
certificate OF mailing (37 CFR 1.8) of Deposit with U.S. Postal Service: December 20, 1991
Penny Drake Name of Person Mailing Paper
IN THE UNITED STATES PATENT AND TRADEMARK OFFICE ,
Art Unit 121
Examiner J. Haley
Applicant(s) Bruce Roth serial Number : 07/660,976
Filed : February 26, 1991
For [R- (R*R*) (4-FLUOROPHENYL)
ACID ITS LACTONE FORM AND SALTS THEREOF Commissioner of Patents and Trademarks Washington, D.C. 20231
NOTICE OF APPEAL under 37 CFR 1.191
Sir:
Applicant(s) hereby appeals to the Board of Appeals the decision of the Primary Examiner twice rejecting finally rejecting claims 4-15 dated September 23, 1991. Applicant(s) further requests that the appeal fee, $ 120.00 , or such greater or lesser amount as the Commissioner may deem required under 37 CFR 1.17(e), be charged to Deposit Account No. 23-0450.
This notice is being filed under 37 CFR 1.8 and the required Certificate of Mailing appears above.
Applicant(s) filed a response to the Examiner's final rejection on , less than two months after the final rejection, and received an advisory action dated , which date constitutes the end of the extended period for response pursuant to 1027 og 71.
23-0450 030 219 130.00CH
PLD- 19 1 Page 1 of 2 5/90 Applicant(s) hereby petitions for a month extension of the period for response in the above-cap ioned application which expired on and hereby authorizes the Commissioner to charge the extension fee of $ , or such greater or lesser amount as may be required by 37 CFR 1.17, to Deposit Account No. 23-0450. This transmittal letter is submitted in triplicate. If for any reason the extension requested above is insufficient to extend the period for filing an appeal under 37 CFR 1.191 to the date of this paper, Applicant(s) hereby petitions for an extension of time under 37 CFR 1.136(a) sufficient to obtain the necessary extension and/or the revival of the above-captioned application as having been unintentionally abandoned and authorizes the Commissioner to charge the required fees under 37 CFR 1.17 to Deposit Account No. 23-0450.
Respectfully submitted,
December 20, 1991 Data Registration No. 25,968 Warner-Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 Tel. No. (313) 996- 7530
Attachments: Form PLD-191 (in triplicate)
PLD- 19 1 Page 2 of 2 5/90 PATENT/Docket
1.. ., certificate OF MAILING (37 CFR Date of Deposit with U.S. Postal Service: January 15, 1992 hereby certify that this transmittal together with the appeal brief referred to below is being deposited with the United States Postal Service as first class mail under 37 CFR 1.8 on the date indicated above and is addressed to the Commissioner of Patents and Trademarks, Washington D.C. 20231.
Penny Drake Name of Person Mailing paper
IN THE UNITED STATES PATENT AND TRADEMARK OFFICE BEFORE THE BOARD OF PATENT APPEALS
Art Unit : 121 Examiner J. HALEY Appellant(s) : BRUCE DAVID ROTH Serial Number : 07/660,976 filed : FEBRUARY 26, 1991 For : 1- METHYLETHYL-3 ACID, ITS LACTONE FORM AND SALTS THEREOF
commissioner of Patents and trademarks Washington , D.C . 2023 1
TRANSMITTALOF APPEAL BRIEF UNDER 37 CFR 1.192
Sir:
Appellant(s) hereby transmit three (3) copies of the brief required under 37 CFR 1.192 in connection with the appeal in the above-captioned application. The notice OF appeal UNDER 37 CFR 1 191 was filed on . December 20, 1991 . appellant(s) hereby request. that the fee for filing a brief in support of an appeal, $ 260.00 , or such greater or lesser amount as the Commissioner may deem is required by 37 CFR 1.17( f), be charged to deposit Account No. 23-0450.
The brief is being filed under 37 CFR 1.8 and the required Certificate of Mailing appears above.
Appellant(s) hereby request an oral hearing pursuant to 37 CFR 1.194 and hereby request that the fee for filing a request fox oral hearing, , or such greater or lesser amount as the Commissioner may deem is required by 37 CFR 1.17(g), be charged to Deposit Account No. 23-0450.
Page 1 of 2 PATENT/Docket No: 3844-C1-01-JT
[ Appellant(s) reserve the right to request an oral hearing pursuant to 37 CFR 1.194 following receipt of the Examiner's Answer.
Appellant(s) hereby petition for a month extension of the period for filing the brief, which expired 2 months from the date of the NOTICE OF APPEAL (or the date of the Examiner's action noted in the N notice OF APPEAL, if later), and hereby authorize the Commissioner to charge the extension fee of $- , or such greater or lesser amount as may be required by 37 CFR 1.17, to Deposit Account No. 23-0450. This transm ttal letter is submitted in triplicate. If for any reason the extension requested above is insufficient to extend the period for filing the brief under 37 CFR 1,192 to the date of this paper, Appellant(s) hereby petition for an extension of time under 37 CFR 1.136(a) sufficient to obtain the necessary extension and/or the. revival of the above-captioned application as having been unintentionally abandoned and authorize the Commissioner to charge the required fees under 37 CFR 1.17 to Deposit Account No. 23-0450.
respectfully submitted,
, Attorney Registration No. 25,968 Warner -Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 Tel. NO. (313) 996-7530
attachments Form pld-192 (in triplicate) AppelIant's brief (In triplicate)
pld 192 Page 2 of 2 11/90 PD-3844-C1-01-JT
lN THE united STATES PATENT & TRADEMARK OFFICE BEFORE THE HONORABLE BOARD OF patent APPEALS AND INTERFERENCES
BRUCE david ROTI-I examinar J. HALEY serial NO: 07/660,976 ART UNIT: 121
FILED: FEBRUARY 26, 1991 PAPER NO: 12
FOR :
ACID, ITS LACTONE form AND SALTS THEREOF
January 15, 1992 APPEAL BRIEF
Hon Commissioner of Patents and Trademarks Washington, D.C. 20231
Dear Sir: This is an appeal of the final rejection of claims 4-15, all. of the pending claims in the application.
STATUS OF CLAIMS
There were 12 original claims in the specification as filed, claims 1-3 were cancelled during the prosecution of the application and new claims ’13-15 were added at the same time. The claims on appeal are claims 4-15, which appear in Appendix A.
status OF AMENDMENTS
There were no amendments made following the final rejection of September 23, 1991 . an interview was held with the Examiner on November 4, 1991, where the final rejection was sustained. 07/660,976 -2- PD-3844-C1-0l-JT
solution of converting to the open ring form, the heptanoic acid. Administration of the lactone form would result in conversion to the ring open form in the body. Both forms are administrable and capable of being prepared into pharmaceutical compositions. The compounds are useful as hypocholesterolemic agents either in the ring opened or lactone forms. The particular optical isomer claimed in this invention is prepared in accordance with the methods specified in the specification and illustrated on pages 7 and 8 as well as particularly exemplified in the Working Examples. In addition, the pharmacological activity of the particular optical isomer claimed in this invention is compared with the prior known mixture of optical isomers and the opposite isomer, the S isomer, on page 8 of Appellant's specification. The R isomer as claimed appears to be at least 100 times more active than its corresponding S isomer and more than 10 times more active than the mixture. Under ordinary circumstances one would have expected only a two-fold difference between the particular R isomer and the mixture.
ISSUE ON APPEAL
Whether the rejection of claims 4-15 under 35 U.S.C. 102(b) as allegedly being anticipated by Roth, U.S. Patent 4,681,893, should be sustained.
REFERENCES
The reference applied in the final rejection is U.S. Patent to Roth 4,681,893 issued July 21, 1987.
grouping OF CLAIMS
Claim 13 is directed to the particular R isomer in either the ring open heptanoic acid form or in the lactone or ring closed form the pyrrole-3 - carboxamide and pharmaceutically acceptable salts. claim 14 specifically claims the ring opened R optical isomer of claim 13. Claim 15 particularly claims the R optical isomer of the ring closed or lactone form of claim 13. Claims 4-10 refer to particular salts of the ring opened cornpound of claim 14. Finally, claims 11 and 12 claim pharmaceutical compositions and method of treatment for hypocholesterolemia emplaying the compound of claim 13 in either the ring open acid form or the lactone form. 07/660 , 976 -3- PD-3844-C1-0l-JT
Since the rejection is on the basis of anticipation, 35 U.S.C. 102(b), claims 4, 5 and 11-15 stand or fall with claim 13. Claim 6-10 should be separately considered as not being disclosed or even hinted at in U.S. Patent 4,681,893.
ARGUMENT
The prior art reference does not anticipate the claims on appeal. in accordance with the well-established principles for determining anticipation.
To anticipate under 35 U.S.C. 102(a), a prior art reference must disclose each and every element of claimed invention, or their equivalence, and the element must function in substantially the same way to produce substantially same result; identity of structure, purpose and result is required to make out anticipation. (TateEngineering, Inc. united States al., 175 USPQ 115)
The prior art reference, U.S. Patent 4,681,893 states the following at column 3, lines 45-54:
The compounds of structural formula I above possess two asymmetric carbon centers, one at the 4-hydroxy position of the pyran-2-one ring, and the other at the 6-position of the pyran-2-one ring where the alkylpyrrole group is attached. This asymmetry gives rise to four possible isomers, two of which are the R-cis- and S-cis-isomers and the other two of which are the R-trans- and S-trans-isomers. This invention contemplates only the trans- form of the compounds of formula I above.
The prior art reference thus mentions the possible existence of four separate isomers. Of those four, the reference identifies two possibilities as being preferred, namely the R- and S-trans- forms. Nowhere does the prior art state or suggest which optical isomer is preferred and does not mention how one skilled in the art could make the pure optical isomers separately. The above paragraph taken out of column 3 of U.S. Patent 4,681,893 is the only reference in the patent to the existence of optical isomers. In contrast, the present invention describes the particular R isomer which is found to have greater than 10 times the activity of the compound described in the prior art reference, namely, the racemic mixture. It therefore becomes clear that the compound of the present invention although having the same general utility, is not identical in actual spatial configuration and does not 07/660,976 - 4- PD-3844-C1-01-JT produce substantially the same result since it has greater than 10 times the activity than the reference compound. It is well established patent law that a prior art reference does not anticipate unless its disclosure is enabling in that it describes in full, clear and exact terms as to enable any person skilled in the art to practice the invention. The mere suggestion of isomers in the Roth patent, U.S. 4,681,893, is not enabling and therefore is not anticipatory. See for example Monsanto company V. Dawson Chemical Company et. al. , 165 USPQ 560 and In re wiggins James, and Gittos, 179 USPQ 421 where the Court stated: "naming of compounds by reference does riot constitute a description of invention within meaning of 35 U.S.C. 102(b).. and also re Donohue, 226 USPQ 619. The Examiner rejected Appellant's claims relying on In re schaumann, 197 USPQ 5. A recent case supporting Schaumann is Ex parteA at 17 USPQ2d 1716. The Board also recognized the need for an enabling disclosure in holding that Applicant's claim for a chemical compound was anticipated by the prior art reference because the prior art reference named that compound among 45 others and there was no dispute that the synthetic procedures disclosed enabled the preparation of the cornpound which Appellant tried to claim. In the present case, the Examiner has held that Schaumann apllies because of the limited number of possible isomers existing and states that "a sufficient description of each jsomer is provided by the reference". Unfortunately, Appellant cannot see such a "sufficient description of each isomer'' in column 3, lines 45-54. The Examiner refers to this passage as an "overt statement of contemplated isomers", Nowhere does this "overt statement" recognize the fact that the R isomer is the most desired and the most surprisingly active isomer of the two possibilities if one is to select from the trans compounds and, furthermore nowhere does it suggest or teach how one can prepare, isolate or separate the mixtures to prepare or isolate the desired R isomer free from the corresponding S isomer. Schaumann stands for- the proposition that there can be anticipation when a reference embraces a very limited number of compounds closely related to one another in structure so that one can arrive at a conclusion that the reference provides a description of those compounds just as surely as if they were identified in the reference by name. In contrast, the present case does not fall within the facts of Schaumann. The present invention is directed to a novel optical isomer which was not made and not known to have the properties it has at the time of 07/660,976 -5- PD-3844-Cl-01-JT
the Roth patent, U.S. 4,681,893. Furthermore, the description in the Roth patent is not such that one skilled in the art would have been possessed of the isomer at that time. The Roth description, as above, merely talks of the possibility of four isomers, two of which are preferred. Nowhere does it mention how to isolate the individual optical isomers to find out which is the most preferred. Schaumann is demonstrably not controlling in this case on analysis of In re May and Eddy, 197 USPQ 601, 607. The facts in Mayand Eddy are similar to that of the present invention in that it concerns optical isomers, racemates and diastereomers. At page 607 of May and Eddy, the novelty of claim 6 directed to the use of a hydrochloride salt was found anticipated by the prior art disclosure of various salts and in particular the hydrobromide salt. The Court relied on Schaumann stating: "a reference does not fail as an anticipation merely because it does not contain a description of the subject matter of the appealed claim in ipsissimis verbis". Nevertheless, when referring to other particular method of use claims, the Court held that these were critically different from claims 1 and 6 in that they recite the use of a novel compound as recognized in In re Williams, 36 CCPA 756, 171 F.2d 319, 80 USPQ 150 (1948), "the novelty of an optical isomer is not negated by the prior art disclosure of its racemate".
CONCLUSION
For reasons gi above, reversal of the rejection under 35 U.S.C. 102(b) of claims 4-15 is respectfully requested. The Examiner's rejection is erroneous as a matter of law by applying the facts of the present case to the wrong law. The issue here is whether an optical isomer is novel over its prior disclosed racemic mixture. The law as stated in May and Eddy affirming In re Williams says yes.
Respectfully submitted ,
Reg. No. 25,968 Warner-Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 (313) 996-7530 RAD:cm/056 Attachment 07/660,976 - 6-
APPENDIX
4. The monosodium salt of the compound of Claim 14.
S. The salt of the compound of Claim 14,
6. The hemicalcium salt of the compound of Claim 14.
7. The N-methylglucamine salt of the compound of Claim 14.
8. The salt of the compound of Claim 14.
9. The hemizinc salt of the compound of Claim 14.
10. The mixture with the compound of Claim 14.
11. A pharmaceutical composition for treating hypercholesterolemia comprising a hypocholesterolemic effective amount of a compound of Claim 13 and a pharmaceutically acceptable carrier.
12. A method of inhibiting cholesterol synthesis in a human suffering from hyperchoiesterolemia comprising administering a of Claim 13 in unit dosage form.
13. -4- acid (1- -N,4 1- 2- (tetrahydro-4-hydroxy-6-oxo- -2H- pyran-2- or pharmaceutically acceptable salts thereof,
14. A compound of Claim 13 which is
acid.
15. A compound of Claim 13 which is -2-- ( 1 -N, 1- 2- -6-oxo- -1H- pyrrole-3-carboxamide. UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office ASSISTANT SECRETARY AND COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 2023 1
BEFORE THE BOARD OF PATENT APPEALS AND INTERFERENCES
Serial Number: 07/660976 Filing Date: 02/26/91 Appelant(s): BRUCE D. ROTH ET AL.
JOAN THIERSTEIN for Appellant
EXAMINER'S ANSWER
This is in response to appellant's brief on appeal filed
February 26, 1991. (1) Status of claims. The statement of the status of claims contained in the brief
is correct.
This appeal involves claims 4-15. No claims are allowed.
(2) Status of Amendments After Final. No amendment after final has been filed.
(3) Summary of invention.
(4) Issues.
The appellant's statement of the issues in the brief is
correct. Serial No. 07/660796 -2-
Art Unit 1201
(5) Grouping of claims. The appellant's statement in the brief that certain claims do not stand or fall together is not agreed with because
appellant urges that none of the specific salts of claims 6-10 "are disclosed or even hinted at in the reference. Such is not
seen to be so since column 7, lines 8-18 disclosesthe claimed metal salts of claims 6, 8 and 9. Also, the amine salt of claims
7 and 10 are generically disclosed.
(6) Claims appealed. The copy of the appealed claims contained in the Appendix to the brief is correct.
(7) Prior Art of record. The following is a listing of the prior art of record relied upon in the rejection of claims under appeal.
4,681,893 Roth 7-1987 (8) New prior art. No new prior art has been applied in this examiner's answer.
(9) Grounds of rejection. The following ground(s) of rejection are applicable to the appealed claims.
Claims 4-15 are rejected under 35 U.S.C. § 102(b) as being
anticipated by Roth, 4,681,893. Serial No. 07/660796 -3- Art Unit 1201
Roth teaches the claimed substituted hydroxypyran-2-ones and their corresponding dihydroxy acids as useful inhibitors of cholesterol synthesis. See col. 2, formula
I and lines 39-43; col. 6 formula x. Regarding the stereochemistry of these compounds, col 3 lines 49-54 restrict the invention to the trans-isomers and col. 6 lines 56-58 specify the R*,R* configuration. The salt forms are disclosed at col. 7, lines 8-18. The pharmaceutical compositions are set forth at col. 8, lines 18-22.
(10) New ground of rejection. This Examiner's Answer does not contain any new ground of rejection.
(11) Response to argument. Appellant urges that the reference does not provide a sufficient description of each individual isomer. Where a reference discloses a genus of compounds of similar structure which are sufficiently limited in number, the reference is deemed to provide description of those compounds just as specifically as if they were identified by name. In re Shaumann, 197 USPQ 5
(CCPA 1978). In the instant case, the Roth reference teaches how to make the racemic mixture. Further, the reference teaches (at column 6, lines 56-59) a preference for the R*,R* isomer. One having ordinary skill in the art would know how to resolve the racemic mixture to attain the preferred isomer. Such knowledge Serial No. 07/660796 -4- Art Unit 1201,
is conventional in the art. Further, even if a preferred isomer were not disclosed, one skilled in the art expects one of the individual isomers to be more active than the other since this, too, is knowledge contemporary in the art. Appellant appears to be urging that one skilled in the art would not possess this knowledge without explicit teaching. However, one skilled in the art is not a novice chemist who is left to blind experimentation. The skilled artisan is just that, a person of ordinary knowledge in the art. Thus, one of ordinary skill is charged with the knowledge common in the art. Therefore, the Roth reference is deemed to place the claimed invention in full possession of the public. For the above reasons, it is believed that the rejections should be sustained. Respectfully submitted,
HALEY :mp March 11, 1992 March 18, 1992
WARNER-LAMBERT COMPANY 2800 PLYMOUTH ROAD ANN ARBOR, MI 48105 PATENT/Docket No:
CERTIFICATE OF MAILING (37 1.8) Date of Deposit with U.S. Postal Service: April 7, 1992 by certify that this paper is being deposited with the United State Service as first class mail under 37 CFR 1.8 on the date indicated above s addressed to the Commissioner of Patents and Trademarks, Washington D.C.
Signature
IN THE UNITED STATES PATENT AND TRADEMARK office Art Unit : 121 Examiner J. HALEY Appellant(s) : BRUCE DAVID ROTH Serial Number : 07/660,976 Filed : FEBRUARY 26, 1991 For PHENYL-4- ACID, ITS LACTONE FORM AND SALTS THEREOF
Commissioner of Patents and Trademarks Washington, D.C. 20231
REQUEST FOR ORAL HEARING UNDER 37 CFR 1.194
Sir:
Appellant(s) hereby request an oral hearing in the above-captioned application and appeal. Appellant(s) further request that the fee for filing a request for oral. hearing, $ 220.00 , or such greater or lesser amount as the Commissioner may deem is required by 37 CFR 1.17(g), be charged to Deposit Account No. 23-0450.
The request for oral hearing is being filed under 37 CFR 1,8 and the required Certificate of Mailing appears above.
The date of the Examiner's Answer in this appeal is March 24, 1992 .
3 Appellant(s) hereby petition for a month extension of the period for filing a request for oral hearing in the above-captioned application which expired one month after the date of the Examiner's Answer indicated above, and hereby authorize the Commissioner to charge the extension fee of , ox such greater or lesser amount as may be required by 37 CFR 1.17, to Deposit Account No, 23-0450.
3/91 PATENT/Docket No: 3844-C1-01-JT
If for any reason the extension requested above is insufficient to extend the period for filing the request for oral hearing under 37 CFR 1.194 to the date of this paper, Appellant(s) hereby petition for an extension of time under 37 CFR 1.136(a) sufficient to obtain the necessary extension and authorize the Commissioner to charge the required fees under 37 CFR 1.17 to Deposit Account No. 23-0450.
Respectfully submitted,
Registration No. 25,968 Warner-Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105 Tel. No. (313) 996-7530
Attachment: [X] Form PLD-194 (in triplicate)
PLD 194 Page 2 of 2 Pages 3/9 1. U.S. DEPARTMENT OF COMMERCE 15 MAILED Patent and Trademark Office ASSISTANT SECRETARY AND COMMISSIONER OF PATENTS AND TRADEMARKS JUL 2 9 1992 Washington, D.C. 20231
PAT. &T.M. OFFICE BOARD OF PATENT APPEALS 15 AND INTERFERENCES PaperNo. 1rc
Joan Thierstein Appeal No: 92-2941 Warner-Lambert Company Appellant: Roth, Bruce D. 2800 Plymouth Road SerialNo.: 07;660,976 Ann Arbor, MI 48105 Hearing Docket: A Date: October 19,1992 Time: 9100 a.m. Place: Room 12C07 CRYSTAL GATEWAY 2 1225 Jefferson Davis Highway Arlington, VA 22202 NOTICE OF HEARING CONFIRMATION REQUIRED WITHIN TWENTY-ONE DAYS
Your attention is directed to 37 CFR 1.194(a). The above identified appeal will be heard by the Board of Patent Appeals and Interferences on the date indicated. Hearings will commence at the time set and as soon as the argument in one appeal is concluded, the succeeding appeal will be taken up.
The time allowed for argument is twenty minutes unless additional time is requested and permitted before the argument is commenced.
CONFIRMATION OR WAIVER OF THE ORAL HEARING IS REQUIRED. The attached form must be filed with the Board within TWENTY-ONE (21) DAYS from the mailing date of this notice, indicating confirmation or waiver of the hearing.
Failure to file the form within the above time period will be construed as a waiver of the request for the oral hearing.
37 CFR 1.136(a) does not apply.
By Order of the Board of Patent Appeals and Interferences.
B.P.A.I. FAX No. Is: (703)557-8642 See 1108 OG 15 (Nov. 14,1989)
Clerk of Board (703) 557-4101
In all communications relating to this appeal, please identify the appeal by its number. D OF PATENT APPEALS AND INTERFERENCES MISSIONER OF PATENTS AND TRADEMARKS washington D.C. 20231
MI 48105 FROM:
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Hearing attendance confirmed.
Hearing waived.
B.P.A.I. FAX No. Is: (703) 557-8642 See 1108 OG 15 (Nov. 14,1989)
(Signature) RONALD A. PATENT COUNSEL WARNER-LAMBERT COMPANY
PTOL-38 5-90) IN THE UNITED STATES PATENT AND TRADEMARK OFFICE
In re application of: B.D. ROTH Serial No.: 07/660,976 Group No.: Filed: FEB. 26, 1991 Examiner: J. HALEY For: [R- (R*R*)
ACID, ITS LACTONE FORM AND SALTS THEREOF
Commissioner of Patents and Trademarks Washington, D.C. 20231
APPEAL NUMBER 92-2941
NOTICE OF HEARING
[ HEARING ATTENDANCE CONFIRMED FOR 9:00 A.M. OCTOBER 19, 1992
CERTIFICATE OF MAILING (37 CFR 1.8a)
I hereby certify that this paper (along with any paper referred to as being attached or enclosed) is being deposited with the United States Postal Service on the date shown below with sufficient postage as first class mail in an velope addressed to the: Commissioner of Patents and Trademarks, Washington, D.C. 20231 PENNY DRAKE (Type or print name of person mailing paper) Date: AUGUST 11, 1992
(Signature of personmailing paper) Art Unit 1201 Paper No. 17 . Appeal No. 92-2941 MTC
MAR 16 1993 HEARD : October 19, 1993
UNITED STATES PATENT AND TRADEMARK OFFICE
BEFORE THE BOARD OF PATENT APPEALS AND INTERFERENCES
Ex parte Bruce D. Roth
Application for Patent filed February 26, 1991, Serial No. 660,976, which is a continuation of Serial No. 384,187, filed July 21, 1989, now abandoned.
Acid, Its Lactone Form And Salts Thereof.
Ronald A. Daignault et al. for Appellant.
Supervisory Primary Examiner - Mary C. Lee. Examiner - J. Haley.
Before Seidleck, Meros and J, Smith, Examiners-in-Chief.
seidleck, Examiner-in-Chief.
This appeal involves claims 4 through 15, all of the
claims in the case. A copy of the appealed claims is attached to this
opinion as an appendix. Appeal No. 92-2941
The examiner has relied upon the following reference in the answer:
Roth 4 , 681,893 July 21, 1987 Claims 4 through 15 stand rejected under 35 USC §102(b) as being anticipated by Roth. We have carefully considered all of the arguments advanced by appellant and the examiner and find that we agree with appellant. Accordingly, we will not sustain the aforementioned rejection. The present invention is directed to compounds consisting of
Acid, pharmaceutically acceptable salts thereof and
(the Lactone Form of the heptanoic acid). As interpreted by both appellant and the examiner throughout prosecution, the claimed subject matter is specifically directed to essentially pure materials, more particularly, the isomer which is essentially free of any of the isomer also present in the trans racemate.
-2- Appeal No. 92-2941
As painted out by the examiner, and as acknowledged by appellant, the Roth reference discloses certain or ones and the ring-opened hydroxy acids derived therefrom. Roth teaches that these trans compounds, because of the asymmetric carbon centers, give rise to both the R-trans and the
S-trans isomers, note column 3, lines 49 to 54, column 6, lines 56 to 58 and example 2. However, as correctly observed by appellant, Roth, at best, only describes the trans racemate containing the R-trans and the S-trans isomers in admixture. Nowhere does Roth state or suggest which optical isomer is preferred and, moreover, does not specifically mention haw one skilled in the art could make the pure optical isomer separately. In view of the above, we are unable to subscribe to the examiner's contention that the Roth disclosure anticipates the claimed subject matter. The pure R-trans isomer is simply not described in the Roth reference within the meaning of
35 USC §102. As recognized in In re Williams, 171 F.2d 319, 80
USPQ 150 (CCPA 1948) and re-affirmed by our reviewing court in In re May, 574 F.2d 1082, 197 USPQ 601, 607 (CCPA the novelty
-3- Appeal No. 92-2941
of an optical isomer is not negated by the prior art disclosure of its racemate. Thus, the disclosure in Roth of the racemate containing both the R-trans and S-trans isomers does not negate the novelty of the presently claimed purified R-trans isomer. The examiner's reliance upon the holding in In re Schaumann, 572 F.2d 312, 197 USPQ 5 (CCPA 1978), in support of the position that the claimed pure R-trans isomer lacks novelty in view of the disclosure in Roth, is misplaced. The court in In re May, supra, recognized that claims directed to optically pure isomers present issues critically different from those presented by the facts in In re Schaumann, supra. Such is evident from the aforementioned specific holding in In re May, supra, that novelty of an optically pure isomer is not negated by the prior art disclosure of its racemate. Upon further prosecution of this application before the examiner, we recommend that the examiner analyze the claimed subject matter under the provisions of S103 of 35 USC. An obviousness rejection of claims directed to an optically pure isomer appears to be in order when, as here, (1) the product of the prior art is known to be racemic and (2) where methods for resolving the racemic mixture into the pure optically active
-4- Appeal No. 92-2941
isomers are known to those skill in the art. In re Williams, supra.
The decision of the examiner is reversed. REVERSED.
James A. Seidleck Examiner-in-Chief
) BOARD OF PATENT APPEALS ) INTERFERENCESAND Examiner-in-Chief
Joan Thierstein Warner-Lambert Company 2800 Plymouth Road Ann Arbor, MI 48105
-5- APPENDIX
4. The monosodium salt of the compound of Claim 14,
5. The monopotassium salt of the compound of Claim 14.
6. The hemicalcium salt of the compound of Claim 14.
7. The N-methylglucamine salt of the compound of Claim 14.
8. The hemimagnesium salt of the compound of Claim 14.
9. The hemizinc salt of the compound of Claim 14.
10. The mixture with the compound of Claim 14.
11. A pharmaceutical composition for treating hypercholesterolemia comprising a hypocholesterolemic effective amount of a compound of Claim 13 and a pharmaceutically acceptable carrier.
12. A method of inhibiting cholesterol synthesis in a human suffering from hypercholesterolemia comprising administering a compound of Claim 13 in unit dosage form.
13,
-1-heptanoic acid or
-2H--- or pharmaceutically acceptable salts thereof.
14. A compound of Claim 13 which is
acid.
15. A compound of Claim 13 which is 2-
pyrrole-3-carboxamide. . UNITED STATESDEPARTMENT OF COMMERCE Patent and Trademark Office Address: COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231 -- NUMBER I FILINGDATE I FIRST NAMED APPLICANT ATTORNEY DOCKET NO. "- "-
NOTICE OF ALLOWABILITY
PART 1. This communication is responsive to 2. All the claims being allowable, PROSECUTION ON THE MERITS remains CLOSED in this application. If not included herewith (or previously mailed), a Notice Of Allowance And issue Fee Due or other appropriate communication will be sent in due course. 3. The allowed claims are I 4. The drawings filed on are acceptable. 5. Acknowledgment is made of the claim for priority under 35 U.S.C. 119. The certified copy has been received. not been received. been filed in parent application Serial No. , flied on 6. Note the attached Examiner's Amendment. 7. Note the attached Examiner Interview Summary Record, PTOL-413. 8. Note the attached Examiner's Statement of Reasons for Allowance, 9. Note the attached NOTICE OF REFERENCES CITED. PTO-892. 10. Note the attached INFORMATION DISCLOSURE CITATION, PTO-1449.
PART II. A SHORTENED STATUTORY PERIOD FOR RESPONSE to comply with the requirements noted below is set to EXPIRE THREE MONTHS FROM THE "DATE MAILED" Indicated on this form. Failure to timely comply will result in the ABANDONMENT of this applicatlon. Extensions of time may be obtained under the provisions of 37 CFR
1. Note the attached EXAMINER'S AMENDMENT or NOTICE OF INFORMAL APPLICATION, PTO-152. which discloses that the oath or declaration is deficient. A SUBSTITUTE OATH OR DECLARATION IS REQUIRED. 2. APPLICANT MUST MAKE THE DRAWING CHANGES INDICATED BELOW IN THE MANNER SET FORTH ON THE REVERSE SIDE OF THIS PAPER. a. Drawing informalities are indicated on the NOTICE RE PATENT DRAWINGS. PTO-948, attached hereto or to Paper No. . CORRECTION IS REQUIRED. b. The proposed drawing correction filed on has been approved by the examiner. CORRECTION IS REQUIRED. c. Approved drawing corrections are described by the examiner in the attached EXAMINER'S AMENDMENT. CORRECTION REQUIRED. d. Formal drawings are now REQUIRED.
Any response to this letter should include in the upper right hand corner, the following information from the NOTICE OF ALLOWANCE AND ISSUE FEE DUE: ISSUE BATCH NUMBER. DATE OF THE NOTICE OF ALLOWANCE, AND SERIAL NUMBER.
Attachments: Examiner's Amendment Notice of InformalApplication. PTO-152 Examlner Interview Summary Record. PTOL- 413 Notice re Patent Drawings, PTO-948 Reasons for Allowance Listing of Bonded Draftsmen . Notice of References Cited, PTO-892 . Other Information Disclosure Citation, PTO-1449
89-3615 UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office Address: Box ISSUE FEE COMMISSIONER OF PATENTS AND TRADEMARK: Washington, D.C.20231
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3. Further correspondence to be mailed to the followlng: 4. For printing on the patent front page, list the names of not more than Ronald A. Daignaul. Elizabeth M. Anderson 3 registered patent attorneys or Warner-Lambert Company agents OR alternatively,the name of a 2 2800 Plymouth Road firm having as a member a registered Ann Arbor, MI 48105 attorney or agent. If no name is listed, no name will be printed. 3
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,- I I (WED)08. 05 98 17:00/No. 3560555427 p
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As discussed yesterday we need to obtain declarations or certifications from the U.S.Patent Office which indicate the expiration date for U.S. Patents 4,681,893;5,273,995 and 5,686,104. These patents arc owned by Warner-Lambert Company. Warner-Lambert filed pipeline applications based on these U.S. patents in brazil The pipeline applications have been granted.
FROM WARNER-LAMBERT CO, (WED)08. 26‘ 98 I4 0 50/ST. 14: 52/N0. 3560555645 P 1
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Francis J. Tinney Phone: (734) 622-7296
VIA FACSIMILE
United States Patent and Trademark Office Attn: Sherry D. Brinkley Petitions Examiner office of Petitions Office of the Deputy Assistant Commissioner For Patent Policy and Projects Washington D.C, 2023 1
In re: Application of U.S.Patent No. 5,273,995 REQUEST FOR Issued: December 28,1993 CORRECTION OF For: [R-(R*R*)]-2-(4-FLUOR0PHENYL)-ß,S- TITLE ON DECISION DIHYDROXY-5-(1-METHYLETHYL- ON COMMUNICATION 3-PHENYL-4-[(PHENYLAMINO)CARBONYL] FILED BY FACSIMILE -1H-PYRROLE-1-HEPTANOIC ACID, ITS ON AUGUST 6,1998 LACTONE FORM AND SALTS THEREOF
Dear Ms, BrinkIey.:
This is a request to correct the titIe on the decision issued pursuant to Applicant's Petition under 37 CFR 1. 182, to certify the date of expiration of the above-identified patent. The title should read as follows: The correct Title is: [R-(R*R*0]-2-(4-FLUOROPHENYL)-ß,8- DIHYDROXY-5-(1-METHYLETHYL- 3-PHENYL-4-[(PHENYLAMINO)CABONYL] -1H-PYRROLE-1-HEPTANOIC ACID, ITS LACTONE FORM AND SALTS THEREOF E FROM WARNER-LAMBERT CO. (WED)08. 26' 98 14.53/ST. 14:52/NO, 3560555615 P 3
USPTO Attn: SherryBrinkley August25, I998 Page Two
- I ---.- 'he Commissioner is authorized to charge any fees, to pay my unpaid fees or pay any grater amount as may be required or credit any overpayment to Deposit Account No, 23-0455.
Please forward the corrected certified decision by Federal Express mail (Account No. 0482- 0660-3) to the attention of:
Francis J. Tinney Warner-Lambert Company PatentDepartment 2800 Plymouth Road Ann Arbor, MI 48105 1 Upon receipt of this request please contact me at the above number and let me know when the corrected decision WilI be forwarded.
Should you have any questions, please feel free to contact me.
Thank you for your assistance.
Senior Cod Pharmaceutical Patents UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office ASSISTANT SECRETARY AND COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231
Paper No. 21
Francis J. Tinney Senior Counsel, Pharmaceutical Patents COPY MAILED WARNER LAMBERT Corporate Legal Division 2800 Plymouth Road AUG 1 9 1998 Ann Arbor, MI 48106 SPECIAL PROGRAMS OFFICE DAC FOR PATENTS In re Application of U.S. Patent No. 5,273,995 Issued: December 28, 1993 ON PETITION For: [R- (R*R*)]-2-(4-FLUOROPHENYL) -BETA, : DELTA-DIHYDROXY-5-(1-METHYLETHYL- : 3-PHENYL-4-[(PHENYLAMINO)CARBONYL] : - 1H-PXRROLE-1-HEPTANO1C ACID, ITS : LACTONE FORM AND SALTS THEREOF
This is a decision on the communication filed by facsimile on August 6, 1998, which is being construed as petitions under 37 CFR 1.182 requesting: (1) a certification of the date of expiration of the above-identified patent; and (2) expedited consideration of the instant petition.
The petitions are GRANTED.
35 U.S.C. §154(a) (2) provides that:
TERM. -Subject to the payment of fees under this title, such grant shall be for a term beginning on the date on which the patent issues and ending 20 years from the date on which the application for the patent was filed in the United States or, if the application contains a specific reference to an earlier filed application or applications under section 120, 121, or 365(c) of this title, from the date on which the earliest such application was filed.
35 U.S.C. §154(c)(1) provides that: DETERMINATION. -The term of a patent that is in force on or that results from an application filed before the date that is 6 months after the date of the enactment of the Uruguay Round Agreements Act shall be the greater of the 20-year term as provided in subsection (a), or 17 years from grant subject to any terminal disclaimers. U.S. Patent No. 5,273,995 Page 2
The papers submitted with the instant petition indicate that the above-identified patent: (1) issued on an application filed on February 26, 1991; (2) does not claim the benefit under 35 U.S.C. §§120, 121, or 365(c) of any application filed earlier than prior application No. 07/384,187, filed July 21, 1989; and is not subject to a disclaimer under 35 U.S.C. §253.
Based upon the information submitted with the instant petition, the above-identified patent will remain in force until December 28, 2010, unless (1) any applicable maintenance fee is not timely paid; (2) the above-identified patent is held invalid or unenforceable during a court action; (3) the above-identified patent is disclaimed, either by a statutory disclaimer of all claims or a terminal disclaimer; (4) all of the claims of the above-identified patent are canceled during a reexamination proceeding; or (5) the above-identified patent is reissued.
Inquiries with regard to this decision should be directed to the undersigned at (703) 305-9285.
Petitions Examiner Office of Petitions Office of the Deputy Assistant Commissioner for Patent Policy and Projects UNITED STATES DEPARTMENT of COMMERCE Patent and Trademark Office ASSISTANT SECRETARY AND COMMlSSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231
Paper No. 22
Francis J. Tinney Senior Counsel, Pharmaceutical Patents WARNER LAMBERT COPY MAILED
In re Application of U.S. Patent No: 5,2737,995 Issued: December 28, 1993 ON PETITION For: (R-R*4*)) -2- (4-FLUOROPHENYL)-BETA, : DELTA-DIHYDROXY-5-(1-METHYLETHYL- : 3-PHENYL-4-( (PHENYLAMIN0)CARBONYL) : -1H-PYRROLE-1-HEPTANOIC ACID, ITS : LACTONE FROM AND SALTS THEREOF
This is a decision on the communication filed by facsimile on August 6, 1998, which is being construed as petitions under 37 CFR 2.182 requesting: (1) a certification of the date of expiration of the above-identified patent: and (2) expedited consideration of the instant petition.
The petitions are GRANTED.
35 U.S.C, §154(a) (2) provides that:
TERM. -Subject to the payment of fees under this title, such grant shall be for aterm beginning on the date on which the patent issuesand ending 20 years from the date on which the application for the patent was filed in the United States or,if the application contains a. specific reference to an earlier filed application or applications under section 1.20, 121, or 365(c) of this title, from the date on which the earliest such application was filed.
35 U.S.C. §154(c) (1) provides that:
DETERMINATION. -The term of a patent that is in force on or , that results from an application filed before the date that is 6 months after the date of the enactment of the Uruguay Round Agreements Act shall be the greater of the 20-year term as provided in subsection (a), or 17 years from grant subject to any terminal disclaimers. Page 2
The papers submitted with the instant petition indicate that the above-identified patent: (1)issued on an application filed on February 26, 1991: (2) does not claim the benefit under 35 U.S.C. §§120, 121, or 365(c) of any application filed earlier than prior application No. 07/384,187, filed July 21, 1989; and is not subject to a disclaimer under 35 U.S.C. §253.
Based upon the information submitted with the instant petition, the above-identified patent will remain in force until December 28, 2010, (1)any applicable fee not timely unless . maintenance is . . paid: (2) theabove identified patent is held invalid or unenforceable during a court action; (3) the above-identified patent is disclaimed, either by a statutory disclaimer of all claims ora terminal disclaimer; (4) all of the claims of the above-identified patent are canceled during a reexamination proceeding; or (5) the above-identified patent is reissued.
Inquiries with regard to this decision should be directed to the undersigned at (703) 305-9285.
'Sherry D. Brinkley Petitions Examiner I office of Petitions Office of the Deputy Assistant Commissioner for Patent Policy and Projects UNITED STATES DEPARTMENT OF COMMERCE Patent and Trademark Office ASSISTANT SECRETARY AND COMMISSIONER OF PATENTS AND TRADEMARKS Washington, D.C. 20231
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:I' Contents
825 Contributors to this issue . 827 Graphical Abstracts . P. Doyle. K.L. Loh, K. M. DeVries 833 Enhancement of stereoselectivity in catalytic cyclopropanation re- odM. S. Chinn actions . Neamati-Mazraeh and S. P. McManus 837 Anchimeric assistance in solvolysis of haloalkylphosphines B. Snider, R. Mohan 84 1 Manganese(lll) based oxidative free-radical cyclizations - II ad S. A. Kates Polycyclization reactions proceeding through tertiary cations Mohan, S. A. Kates, 84 5 Manganese(lll) based oxidative free-radical cyclizations - III . A, Dombroski and B. B. Snider Polycyclization reactions proceeding through secondary radicals P. Fei and T. H. Chan 849 Stereoselective sythesis of a-d-glucopyranosides via 6-silyt-a-D- qlucopyranosyl bromides J. Shields and G. B. Schuster 853 Deoxygenation of epoxides by fluorenylidene H. Buist, H. G. Dallmann, 857 Highly chemo- regio-. and stereoselective introduction of a cis-double T. Rymerson and P. M. Seigel bond into a thia-analogue of stearic acid P. Cho and R. G. Harvey 86 1 Synthesis of polycyclic aromatic fluoranthenes Aguilar, A, Reyes. J, Tamariz 865 Regioselectivity of Diets-Alder additions of 1-acetylvinyl arene- d J.L. Birbaum carboxylates R. Williams and F. D. Klingler 869 Stereoselective allylation for preparation of t-hexose derivatives M. Moriarty. R. Penmasta 873 Synthesis of 1.4-diketones by the coupling reaction of trimethylsilylenol d I. Prakash ethers with lead tetraacetate a77 Novel penlafluorophenyl hypervalent iodine reagents
I. Kolodiazhnyi 88 1 Thiocetenes phospnores Dhimane, J.C. Pommelet. 885 Cyclization of aminomethyleneketenes by intramolecular displacement Chuche. G. Lhommet of chlorine synthesis of bicyclic enaminoesters d M. Haddad
[Continued on outside back cover
Pergamon Press: Oxford New York Beijing Frankfurt Sao Paulo Sydney Tokyo Toronto
I od B) is le.The llate11 catalyst %) by 11can SYNTHESIS OF AN HMG-COAREDUCTASE INHIBITOR; MPA(2 A DIASTEREOSELECTIVE ALDOL APPROACH . one-pot J. E. Lynch*.R. P. Volante,R. V. Wattley,I. Shinkai ormation MerckSharp and DohmeResearch Laboratories 39 and P.O. Box 2000, Rahway,New Jersey 07065
cording ABSTRACT eating to Ilion and Thesynthesis of the ß-hydroxy-6-lactonemoiety of an HMG-CoA reductase inhibitor in Its correct absoluteconfiguration(94%ee) has been accomplished nly in 90 via a diastereoselective aldol reaction between aldehyde 4 and the magnesium ils ofthe enolate ofS(+)1,2,2-triphenylethylacetate, Clai sen condensation, and hydroxyl directed reduction ,of the resulting 8-hyroxy-ß-ketoester.
le would The discovery of mevinolin andcompactin 1b 2 as potent inhibitorsof 3-hydroxy- 3-methylglutaryl coenzyme A reductase has resulted in an intensive effort to develop 3 4 efficient laboratory syntheses of these and other related hypocholesterolemicagents. tron The key structural feature commonto these HMG-CoAinhibitors is the9-hydroxy-8-lactone moiety.5 Furthermore,it appears that this unit in its correct absolute configuration .1986. 5 6 (that which corresponds to and 1b) is essential for enzyme inhibition. Thus.we6,
B,Yang.
V.
7 as well as others ,have been interested in the development of an efficient method for es. for the synthesis of such a structural unit. eri,A Prior syntheses of ß-hydroxyl-6-lactone HMG-CoA reductase inhibitors have been accomplished by initial generation of an appropriate, non-racemic lactone "synthon" 41. followed by the coupling of this unit to the remaining portion of the molecule at a site removed from the asymmetric centers.6 These routes have typically suffered from either 26,1699 inefficient generation of the asymmetric synthon and/or inefficient coupling reactions.
1385 Our strategy is based on the use of a didstereoselective aldol reaction-followedby a Claisen condensation for the generation of the 5(S) stereocenter of keto-alcohol 3 from Acknowledgeme _ ments and Mr aldehyde 4. The 5(S)hydroxylfunction of 3would then be expected to direct the reduc- ma nu scri p t p tion of the 3-keto to the desired 3(R) stereochemistry of 2. group _
1.Alberts J.; Lop Stapley Liesch, 2. Endo, A Me Me 3. Compact 2 34 1983, 1 A long standing problem In the preparation of simple (a-unsubstituted)hydroxy N.L.T Grieco. esters (acids) has been the lack of aldehyde facial selectivity in the aldol reaction Mevinol with chiral acetateenolates8. However, several solutions to this problem have recently 4. Lee, T appeared.' We wish to report here the use of one of these methods for the diastereo- C.H.; selective synthesis of the biphenyl HMG-CoA inhibitor 2. 10 S.; Fa _ 1985. The magnesium(II)enolateof S(+)-2-acetoxy-1,1,2-triphenylethanol(5) (1.1equiv) 5.Stokkei was condensed with aldehyde _4 in tetrahydrofuran at -78" via the procedure of Braun and 9 A.A.; Devant9 to produce the diastereomers6 in 93%yield (SS:SR=97:3, as determinedby PMR Willard and HPLC analysis13.Transesterification of 6 with 1.05 equivalent of sodium methoxide 6. Sletzir in methanol gave methyl ester 7(S/R=97:3) in 95%,yield14.Treatment of ester 7 with _ E.G.; I 3 equivalents of lithio-t-butyl acetate in tetrahydrofuran at -30 to -4O°Cfor 1 hr. 7.Prugh, resulted in theformation of ß-keto-8-hydroxy ester 3 in 90% yield15. Highly stereo- J.R.j specific reduction of the 8-keto functionality was effected using sodium Chem. 2435 borohydride-triethylborane in tetrahydrofuran-methanol (4:l) at -78ºgiving diol _8 io 1984. 93% yield6,11,12.The diol ester was saponified with sodium hydroxide in aqueous Lett. Tetrah methanol, acidified to pH 3.8, and the resulting acid9,waslactonizedby _ heating in toluene at 90" for 8 hr to afford lactone 2 in 85% yield (3R5S:3S5R =97:3 as determined 8. Morris _ Washin by HPLC analysis of the corresponding R(+)-a-methylbenzylamide derivatives)13. The ed. J. enantiomeric purity of2 could be enhanced to >99%ee by recrystallization of the Nelson R(+)-a-methylbenzylamine salt of the acid 9 prior tolactonization14. C,H., New Yo
9. Iwasaw Tetrah M.;F Hagiwa E.; OU Knopfe 10.G.E. Gilfil Novel 1 29, 17 _3 _ va om Acknowledgement: We wish to thank Mr. R. Reamer and Ms. L. DiMichelefor NMR measure- ments and Mr. K. Ryan and Mr.E.Corley for chemicalassistance,and Ms. Lee Dolan for uc- manuscriptpreparation.
1. Alberts, A.O.W., Chen, J.; Kuron,G.;Hunt,V.; Huff, J.; Hoffman. C.;Rothrock, J.; Lopez, M.; Joshua, H.; Harris. E.; Patchett, A.; Monaghan,R.;Currie,S.; Stapley,E.; Albers-Schonberg, G.;Hensens.0.;Hirshfield,J.; Hoogsteen,K.; Liesch, J.; Springer, J. Proc. Natl. Acad.Sci. USA 1980, 77,3957. 2. Endo,A.;Kuroda, M.; Tsujita, Y.; J. Antibiot. 1976, 29, 1346. 3. Compactin-Hsu.C.-T.;Wang. N.-Y.;Latimer,L.H.;Sih,C.J. J, Am. Chem. Soc., 1983, 105, 593. Hirama,M.;Uei,M.ibid,1982. 104, 4251.Girota, N.N.;Wendler, Oxy N.L. TetrahedronLett. 1982.5501. Girota,N.N.;Wendler,N.L.ibid 1983, 3687. Grieco, P.A.; Zelle,R.E.;Lis,R.; Finn,J.J.Am.Chem. Soc.1983,105,1403. ion Mevinolin-Hirama,M.;Iwashita,M,Tetrahedron Lett.1983;1811. tly 4. Lee,T.-J.;Holtz,W.J.;Smith, R.L. J. Org. Chem. 1982, 47, 4750. Kuo, eo- C.H.; Patchett. A.A.; Wendler, N.L.ibid1983,48,1991,Yang.Y.L.;Manna, S.; Falck, J.R.. Yang, Y.-L.Tetrahedron Lett.1984,3563;T.-J. Lee, ibid 1985. 4995. riv) and 5. Stokker.G.E.; Hoffman,W.F.;Alberts. A.W.;Cragoe,Jr., E.J.; Deana, A.A.;Gilfillan, J.L.;Huff. J.W.;Novello, F.C.;Prugh,J.O.; Smith, R.L.; PMR Willard, A.K., J. Med.Chem.1985,28, 347. ide 6. Sletzinger,M.; Verhoeven,T.R., Volante, R.P.; McNamara,N.M.; Corley. with E.G.;Liu,T.M.H.,Tetrahedron Lett. 1985. 2951. hr. 7. Prugh,J.D.;Deana,A.A.. Tetrahedron Lett. 1982, 281. Yang, Y.-L.; Falck, .eo- J.R. ibid1982.4305. Danishefsky, S.; Kerwin,J.R.; Kobayshi, S. J.Amer, ium Chem.Soc.1982,104,358.Prasad, U.; Repic, 0. Tetrahedron Lett. 1984, 243 5 and2435 3391. Rosen,T.;Taschner, M.J.; Heathcock, C.H. J. Org.Chem. in 1984, 49, 3994. Majewski,M.; Clive,D.L.J.; Anderson, P.C.,Tetrahedron ous Lett. 1984,2106. Guinden,Y.; Yoakim,C.;Bernstein.M.A.; Morton,H.E. Tetrahedron Lett. 1985. 1185. in ned 8. Morrison, J.D.and Mosher, H.S. "AsymmetricOrganic Reactions" Am. Chem. Soc.; WashingtonD.C.1976.p.142-152. Soladie,G. in "AsymmetricSynthesis: Vol.2. The ed. J.D. Morrison. 1983,AcademicPress.New York.N.Y.,pp-188-199. Evans, D.A.; the Nelson, J.V.; Taber,T.R.in Topics in Stereochemistry, 1982, 13, 1. Heathcock, C.H.,”AsymmetricInSynthesis"Vol,3..ed.J.D. Morrison.1984, Academic Press, New York, N.Y.,pp174-190. 9. Iwasawa, N.; Mukaiyama,T.Chem.Lett., 1983, 297.Braun,M.;Devant, R. Tetrahedron Lett.1984, 5031.Nagao,Y.;Yamada,S.; Kumagai,T.; Ochiai, M.;Fujita,E. J. Chem.Soc.Chem,Comm.;20,1418 (1984).Nagao,Y.; Hagiwara,Y.; Kumagai,T.;Ochiai, M.;Inoue,T.;Hashimoto, K.; Fujita, E.; J, Org. Chem.; 51. 2393 (1986). Helmchen.G.; Leifkauf, U.; Tauter Knopfel.I., Angew.Chem.Int.Ed.Ingl.;24, 874 (1985). 10. G.E. Stokker, A.W. Alberts. P.S. Anderson, E.J. Cragoe,Jr.,A.A.Deana, J.L. Gilfillan, J. Hirshfield,W.J. Holtz, W.F. Hoffman, J.W. Huff, T.J, Lee, F.C. Novello,J.D.Pruch,C.S.Rooney,R.L.Smith, and A.K. Willard, J.Med.Chem.1986,
_29, 170. 1388 Tetrahedron Printed In G
11. Narasaka, K.; Pai, F.C.; Tetrahedron, 1984, 2233. 12.The ratio syn ofdioltoantiwas98:2. 13. Enantiomeric and diastereomericpurity of 9 was obtained by conversionto the corresponding R(+)-a-methylbenzylamides00 equivalents of 99.9% pure 1 R(+)=a-methylbenzylamineat90ºfor 12 hr) and HPLC separationon Dupont Zorba x silica gel eluting with hex a ne :diethylether:methyl-t-buty1et her : aceticacid (4:3:6:1). 14.The stereochemical integrity of methyl ester 7 was determined by proton magnetic resonance in the presence of thechiral solvating agent L-phenyltrifluoromethylcarbinoland/or Eu(HFC)3.
15. Although the Claisen condensation could be perfomeddirectlyon 6 giving 3 in 85% yield, the higher temperature required for this reaction(25ºC) resulted in epimerizationof the newly generated chiral center. 16. Satisfactory infrared and proton magneticresonance spectra were obtained for all compounds. CHFNanalysiswasobtainedforcompounds2,10.4, and -- 9. (Received in USA 26 December 1986)
La synth l'action de 1 1 dicarbonyl 11- 12 imidates Dans ce par condensat Les résu reactions2+ quatre produi nes-1,515 (4, benzimidazole Cependar pendant quatr excedant 60% La detet de leurs cari references 15. àpriori, par Soit par une dans les con et le xylene sition therm 3844
Journal of Medicinal Chemistry
Volume 28. 1985
JMCMAR 28(1-12) 1-1996 (1985) ISSN0022-2623
PHILIP S. PORTOGHESE, Editor
Patrick E. Hanna, Rodney L. Johnson, and Herbert T. Nagasawa, Senior Editors John L. Neumeyer, Book ReviewEditor
EDITORIAL ADVISORY BOARD. Richard F. Borch William F. Michne Michael J. Rance William T. Comer Richard J. Miller William A. Remers Eric De Clercq J. Lars G. Nilsson David C. Remy A. J. Hopfinger MiguelA.Ondetti Daniel H. Rich PovlKrogsgaard-Larsen Helen H. Ong Svante Wold Ex officio: Paul Anderson
AMERICAN CHEMICAL SOCIETY, BOOKS AND JOURNALS DIVISION D. H. Michael Bowen, Director
Charles R Bertsch,Head, Journals Department Marianne Brogan, Associate Head Mary E. Scanlan,Assistant Manager Anne O'Melia, Assistant Editor Mary L. Orosz,Assistant Editor
Elmer M. Pusey, Jr., Head, Production Department Lorrin R. Garson, Head, Research and Development Department ! J. Med. Chem. 1985,28, 347--358 3-Hydroxy-3-methylglutaryl1-coenzyme A Reductase Inhibitors. 1. Structural Modification of 5-Substituted 3,5-Dihydroxypentanoic Acids and Their Lactone Derivatives
G.E.Stokker,*†W.F.Hoffman,*†A.W.Alberts,‡E.J. Cragoe, Jr.,A.†Deana,†J.L.Gilfillan,‡ J. W. Huff,' F. C. Novello, J. D. Prugh, R. L. Smith,' and A. K. Willard Merck Sharp & Dohme Research Laboratories, West Point, Pennsylvania19486, and Rahway, New Jersey 07065. Received June 21, 1984
A seriesof5-substituted 3.5-dihydroxypentanoic acidsandtheirderivativeshave been prepared and tested for inhibition of HMG-CoAreductase in vitro. In general, unlesscarboxylateaanioncanbeformedand the hydroxy group remain unsubstituted in an erythro relationship, inhibitory activity is greatly reduced. Furthermore, only one enantiomer of the ring-open4 form of lactone 6a(±)posessesthe activity displayed by the racemate. Insertion of a bridging unit otherthanethylor(E)-ethenylbetweenthe5-carbinolmoietyand an appropriate lipophilic moiety(e.g., 2,4-dichlorophenyl) attenuates activity.
Formation of the atheromatous plaque or atheroma is acid 1the dihydroxy acid form of mevinolin, is the most accompanied by the localized deposition of plasma lipids, potent HMG-CoA reductase inhibitor (K, 0.6 nM) re- primarily cholesterylesters,inthe intima of the arterial ported to date! Of even greater interest are the findings wall1,2. Growth of the atheroma eventually leads to con- that compactin11,12and mevinolin9,13are highly effective striction of the coronary arterial lumen and ultimately hypocholeaterolemicagents in several animal species and results in atheroscletosis and coronary heart disease (CH- man. Subsequent to the first reports disclosing the D), the major cause of death and disability in Western structure14andbiologicalactivity8ofcompactin,aseries countries.Theseobservations,coupledwith the compelling of studies directedtoward the development of structurally epidemiologicalevidence implicating hypercholesterolemia simplified HMG-CoA reductase inhibitors were initiated as a primary risk factor for CHD3,4 have stimulated re- in theseLaboratories. Described in this paper are the search on the development of therapeutic agents for pre- resultsof our initial study,15which to delineate key venting and treating atherosclerosis based on the attenu- ation of plasma cholesterol levels! The results of the recently completed Lipid Research Clinics Coronary (1)Ross,R.Annu.Rev.Med.1979.30.1. Primary Prevention Trial (LRC-CPPT)6provide strong support for the bash of this approach. The LRC-CPPT (2)Fuster,V.Scand.J. Haematol.1981,27 (Suppl.38),1. (3) US. Department of Health and Human Services,National clearly demonstrated that reduction of lowdensity lipo- InstitutesofHealth,NationalHeart,Lung,and BloodInsti- protein cholesterol (LDL-C) through dietarymodification tute,“Arteriosclerosis”,Vol.1 and 2, NIH Publiation No. and treatment with the bile acid sequesterant cholestyr- 81-2034 and81-2035, US.G.P.O., Washington,DC,1981. amine, either alone or in combination, diminished the (4)Hamburg,D.A;Elliott,G.R.Arteriosclerosis1982.2,357. incidence of CHD morbidity and mortality in hypercho (5)Prugh, J.D;Rooney,C.S.;Smith,R.L.Annu.Rep.Med. lesterolemicmenhighatrisk for CHD. Nevertheless, the Chem,1983, 18, 161. reduction ofdietarycholesterolandsaturatedfatintake (6)(a)LRC-CPPT,J.Am.Med.Assoc.1984, 251, 351. (b) LRC and the use of bile acid sequesterants often fail to lower CPPT, Ibid. 1984, 251, 365. elevated plasma LDC-C levels to the desired extent, par- (7) Havel,R.J.;Kane,J.P.Annu.Rev. Med.1982.33,417. ticularly in patients with familial hypercholesterolemia (8) Endo,A.;Kuroda,M;Tsujita,Y.J. Antibiot. 1976 29,1346. (FH)7. (9)Alberts,A.W.;ChenJ.;Kuron,G.;Hunt,V.; Huff,J.; Hoff- An attractive and potentially more efficacious way to man, C.;Rothrock,J.; Lopez, M.; Joshua,Harris,H.;E; lowerplasmacholesterollevelswould be to control de novo Patchett,A;Monaghan,R; Currie. S. ; Stapley , E; Albers- cholesterogenesis by selectively inhibiting an early bio- Schonberg, G.; Hensens,O.;Hirshfield,J.;Hoogsteen,K.; synthetic step. The functionalized fungal Liesch,J;Springer,J.Proc.Natl.Acad.Sci.USA.1980.77, highly metab- 3957, olites compactin (ML-236B,CS-500)8andmevinolin (10)Rodwell,V.W.; Nordstrom,J.L;Mitschelen,J. J. Adv. Lipid (MK-803)9arepotentinhibitorsofcholesterolbiosynthesis Res. 1976,14,1. at the level of the major rate-limiting enzyme 3-hydroxy- (11) (a)Tsujita,Y;Kuroda,M.;Tanzawa,K;Kitano.N;Endo,A. 3-methylglutaryl- coenzyme A reductase [HMG-CoA re- Atherosclerosis(Shannon,Irel.)1978,32,307. (b) Kuroda, M, ductase; mevalonate: NADP+ oxidoreductase (CoA acy- Tsujita,Y; Tanzawa,K.;Endo. A. Lipids 1979,14,585. lating), EC 1.1.1.34],l0 which catalyzes the conversion of (12)(a) Yamamoto,A.;Sudo,H.;Endo.A.Atherosclerosis(Shan- HMGCoA to mevalonic acid (eq 1). Indeed, mevinolinic non,Irel.)1980,35,259. (b)Mabuchi,H.;Haba,T;Tatami, R; Miyamoto, S.Sakai,;Y.;Wakasugi,T.;Watanabe,A; Koizumi, J; Takeda, R. N.Engl. J.Med,1981,305,478. (13) (a) Tobert, J. b; Hitzenberger, G.; Kukovetz. W.R .;Holmes, HMG-CoANADPH reductose I. B.; Jones. K. H. Atherosclerosis(Shannon, Irel.)1982, 41, 61. (b) Tobert, d. A; Bell, G. D.; Birtwell, J.; James,I.; Ku- I mevalonic acid SCoA kovatz, W. R;Pryor, J.S.; Buntinx, A.; Holmes, L B.; Chao, Y.S. Bolognese J. A. J. Clin. Invest.1982,69,913. HMG-CoA (14)The X-ray crystal of compactinwasfirstreportedby Brown, A. G.; Smale,T.C.; King, T. J,; Hansenkamp, R; Thompson, R.H.J. Chem.Soc,PerkinTrans.11976,1165. †MerckSharp & Dohme, West Point, PA. Note that therelative configuration in Figure 1 of thecited †MerckSharp & Dohme,Rahway,NJ. referencedoes not agree with the crystal coordinates;we Presentaddress: StuartPharmaceuticals, Division of ICI presenthere the relativeand absolutestereochemical Americas,Wilmington, DE 19897. configuration of compactin 348 Journal of Medicinal Chemistry, 1985, Vol. 28, No. 3 Stokker et SARs for compactin-like mimics and afforded a series of slightly modified procedure of Noltes et al.17 provided moderately effective HMG-CoAreductase inhibitors typ- 4-hydroxy ketone 11 in high yield. Acylation of 11 with itfied by the ring-opened form of lactone 6a 2-bromoacetyl bromide in the presence of pyridine fur- HQ nished bromo acetate 12 which was ring closed to 4- CO2H hydroxy-4-methyl lactone 13 via an intramolecular Re- formatsky reaction.18 Substitution of triethylamine for pyridine in the acylation step resulted in elimination of the 2-bromoacetoxy moiety and isolation of the resultant dienone. An alternate route to lactone 13 starting from 4-hydroxy ketone 11 was investigated. This route involved sequential compactin. R-H acylation, intermolecular Reformatsky reaction with ethyl mevinolinR CH3 2-bromoacetate, basic hydrolysis, acidification. and lac- tonization. This route was abandoned in favor of the more efficient two-step route (vide supra). Ethylene lactone 13a was reduced in the same manner as 6a to provide the corresponding ethyl-bridged compound 16. Refluxing a solution of 13a and toluene in the presence of PTSA (trace) resulted in the smooth conversion of 13a to 17. 5-Methoxy-3-hydroxyheptenoic acid 20 was prepared by treating the dimethyl acetal of 2 with diketene (1 equiv) in the presence of TiCl4 by using the general procedure Cl of Izawa and Mukaiyama19 followed by borohydride re- 6a duction, basic hydrolysis, and acidification of the resultant 3-keto ester 19 as shown in Scheme 111. A similar con- Chemistry. The compounds prepared for this study densation of aldehyde 2 provided a mixture of 3-keto ester are listed in Tables I-III Their syntheses from the 3 and dihydro lactone 4 and, thus, was a less expeditious corresponding aldehydes, exemplified by 2, are shown in route to target lactone 6 than was the dianion procedure Schemes I-III. Condensation of aldehyde 2 with the (Scheme I dianion of ethyl acetoacetate16 followed by borohydride The previously undescribed requisite aldehydes were reduction, basic hydrolysis, acidification, and azeotropic prepared as shown in Schemes IV-VIII. The synthesis removal of water provided a mixture of the trans (6a of the unsaturated aldehyde 55a needed for elaborating and cis (6blactones which subsequently was separated lactone 55 is shown in Scheme IVwith phenanthrene-4- by chromatography (Scheme IThe use of MeOH in the carboxaldehyde as starting material. This procedure was borohydride reduction step was found to be advantageous; also used to prepare the known unsaturated aldehyde replacement of MeOH by EtOH produced some of the precursors for lactones 56-58 corresponding ethyl ester which was more resistant to The 3-(decahydronaphthyl)propanals 25 and 27 (pre- hydrolysis. The resolution of 6awas accomplished via cursors to 51 and 52) were elaborated from 21 as shown formation and chromatographic separation of the diaste- in Scheme V. After high-pressure hydrogenation of 22, the reomeric (R)-a-methylbenzylaminesfollowed by basic acid 23 was converted to aldehyde 25 by the Burgstahler hydrolysis and relactonization to yield 6a and 6a(-). modification of the Rosenmund reduction.20 The ethyl Hydrolysis and acidification of 5-hydroxy-3-keto ester ester of 23 (24, isolated in about an equal amount during 3 without prior reduction resulted in spontaneous lactones the workup of 23) was isomerized with AIC13 (2 equiv) at ization to enol lactone 4. Numerous attempts to reduce room temperature and subsequently hydrolyzed to acid 4 to hydroxy lactone 6, either catalytically or via metal 26, which was converted to aldehyde 27 in the same hydrides, were unsuccessful, Treatment of lactone 6a manner used for 23 25. with NH, provided the erythro amide 10. Catalytic re- The Dibal reduction of nitriles 29,32, and 34 (Scheme duction of lactone 6a provided compound 7 containing VI) provided the aldehydes requisite for preparing lactones a saturated bridging unit. The lactol ethers 8 and 9 were 47, 48, and 53, respectively. Aldehyde 37 (precursor to prepared by diisobutylaluminum hydride (Dibal) reduction 44)21 was prepared by alkylation of phenol 36 with the of 6a followed by treatment with MeOH in the pres- diethyl acetal of 2-bromoacetaldehyde followed by hy- ence of pyridinium p-toluenesulfonate (PPTS). drolysis (Scheme VII). The syntheses of methyl lactones 13, 16, and 17 are Finally, conversion of cinnamaldehyde 2 to propargyl- illustrated in Scheme 11. A tin-mediated aldol condensa- aldehyde 41 (precursor to 42) was effected via the four step tion of aldehyde 2 with 2-acetoxypropene following a sequence 2 38 - 39 40 - 41 by using the general method of Allen and Edens22 as shown in Scheme VIII (15) During the course of this study, a series of mevalonolactone derivatives of the general structure were reported to inhibit (17) Noltes J. G.; Verbeek F.; Creemers, H. M.J. C. Organometal CH3 Chem. Synth. 1970-1971, I 57. In the present case. tri- butylstannyl)ascetone was prepared in situ. (18) For a general method, see Maruoka K.; Hashimoto S.; Ki- tagawa Y.Yamamoto H.;Nozaki H. J. Am. Chem. Soc. 1977 99,7705. (19) Izawa T.;Mukaiyama T. Chem. Left. 1975, 161. (20) Burgstahler, A. W.; Weigel. L. Shaefer, C. G. Synthesis 1976.767. HMG-CoA reductase by Sato et al.: Sato, A.; Ogiso A.: No- (21) During the course of this investigation the 4R.6R dechlom , . . from as possible HMG-CoA Reductase Inhibitors Journal of Medicinal Chemistry, 1985. Vol. 28 No. 3 349
Table 1. Effects of Lactone Modification and Stereochemistry Cl
recryst yield, concn. % no. R solvent % mp, °C formula' µg/mL inhib EtOAc 52 168-169 2 0 10 12 50 15 acetone/ hexane 33 148-150 1 13 5 41 10 64 20 84 acetone/ hexane 17 115-1 17 2 4 5 6 10 27 20 17 50 18 n-BuCI 25 114-1 16 1 26 2 45 5 66 10 72 20 86 n-BuCI 44 114-116 1 0 2 0 4 0 8 0 chromat. acetone/CH2Cl2 33 wax 2 0 4 2 6 4 chromat acetone/CH2Cl2 64 88-93 2 0 4 0 8 6 10 0 acetone/ hexane 63 117-118 1 0 2 0 4 0 8 0 n-BuCl 35 136-138 1 11 2 21 4 36 10 61 n-BuCl/hexane 5.4 135-137 2 3 4 7 10 5 25 15 chromat CHCl3/MeOH 90 108-110 2 0 4 3 10 3 25 5 chromat. CH2Cl2/HOAc 42h gum 1 0 2 0 5 0 10 0 'Analytical results are within 0.4% of the theoretical values unless otehrwise noted See Experimental Section for protocol. Tested in the form indicated since carbooxylate anion could not be formed under this testing protocol. pK, 5.22 (30% EtOH). When tested in the lactone form only 25% inhibition was observed at 50 µg/mL Yield from 12. About a 4:3 ratio of erythro and threo. Overall yield from 18. Anal Calcd: C, 52.68. Found: C, 52.02
The synthesis of 43 (the Z isomer of 6a was accom- hydroxycarborylate forms of each compound were tested accomplished by the catalytic hydrogenation (Lindlar) of 42. for intrinsic inhibitory activity. In each instance, the so- Biological Results and Discussion dium dihydroxycarboxlate form proved more active than the lactone form (see Table I, footnote e). Accordingly, The compounds listed in I-IV were evaluatedfor subsequent tests were done exclusively on the sodium their ability to inhibit solubilized, partially purified rat dihydroxycarboxlate forms unless noted otherwise liver HMG-CoA reductase During the initial phase of this The contributions of lactone moiety Stereochemistry and study, both the lactone and the ring-opened sodiumdi- functionality to intrinsicinhibitory compound 6 are illustrated in Table I. Separation of the lactone (22) Allen C. F H.;Edens C. 0.. Jr. Organic Synthese Wiley mixture 6 into the racemic cis 6band trans 6a New york 1955; Collect Vol. III isomers showed that activity resided principally in the I=
350 Journal of Medicinal Chemistry, 1985, Vol. 28, No. 3 Stokker et al.
Table II Effects of Bridge Modification recryst yield. concn, % no. A solvent % mp, °C formula µg/mL inhibb
n-BuCI 60 96-98 1 29 2 38 5 57 10 80 16 chromat. CH2/CL/acetone 50 87-88 1 14 2 26 4 37 a 63 42 c=c chromat. CH2/CL/acetone 11d viscous oil 0.625 0 3.125 8 chromat. IPA/hexane 50 95-98 43 c=c 44 chromat. CH2/CL/acetone 4.2 oil
45 n-BuC1 13d 133-136
5 16 10 9 20 7 18 19 29
analytical results are within 0.4% of the theoretical values unless otherwise noted see Experimental Section for protocol Equatorial 4-me in lactone by reduction of 13a overall yield from aldehyde. HPLC purification on Dupont silica 10/30 with i- PrOH-hexane (1:19 v/v) at 2 mL/min. Times of elution are 13.2 min for 42 and 21.8 min for 43.
racemic trans lactone 6a Resolution of 6a af- methyl ether (20). These results demonstrate the im- forded enantiomers 6a and 6a their evaluation portant contributions of the carboxylate and 5-hydroxyl showed that the activity displayed by the racemate re- groups to activity. Finally, it should be noted that lactol sulted solely from the dextrorotatory isomer. The addition ethers 8 and 9 displayed greatly diminished activities. of a methyl group to the 4-position of 6ato give trans The effects of altering the moiety bridging the aromatic lactone 13a a compound which more closely resembles the and the lactone fragments in 6aon intrinsic inhibitory HMG moiety of the substrate HMG-CoA, did not alter activity are shown in Table II Saturation of the ethenyl activity appreciably. However, cis lactone 13b which bridge in 6a and ita 4-methylderivative 13agave possesses the opposite relative stereochemistry at C-4, was ethyl-bridged compounds 7 and 16, respectively, with little much less active as anticipated from the results obtained change in activity. However, other modifications of the for 6a and 6b Interestingly, oxidation of the 4- bridge such as replacement with the ethynyl (42 cis- hydroxyl group of 6ato provide enol 4 greatly reduced ethenyl(43), and oxymethylene 44groups resulted in loss activity. This result is a likely consequence of the fact that of activity as did complete removal 45 of the bridging 4 readily forms the sodium salt of the enolate and, moiety. In a companion series of naphthalene analogues therefore, fails to undergo ring opening to afford the re- (Tabla II compound 47 containing the saturated two- quired carboxylate anion under alkaline conditions Re- carbon bridge proved superior. Increasing the length of placement of the enolic hydroxyl group in 4 with a methyl the bridge to three carbons (48) reduced activity and group to provide 17 further reduced activity. Replacement elimination of the bridge to provide 46 further reduced of the carboxyl group in the ring-opened form of 6a activity. with a carboxamido group 10 ablated activity as did The results of various carbocyclic moieties substituted Table 111. Effects of 6-Substitution
xyield.' recryst concn. % no. X solvent % mp, °C formula µg/mL inhib 49 Et2O//pet ether ~8-10 56-58 5 1 10 0 20 6 50 36 50d Et2O/ hexane ~8-10 69-70.5 5 23 10 6 20 17 50 34 51 Et2O/ hexane ~8-10 68-71 5 15 10 22 20 44 50 70 52' Et2O/ hexane ~8-10 102-110 1 1 5 13 12.5 28 53 H Et2O/ hexane ~8-10 122-129 1 18 5 16 12.5 28 54 Et2O 23 96-98 5 3 10 15 20 14 50 31 55 n-BuC1 25 140-142 0.35 39 0.75 58 1.0 64
56h n-BuC1 3 151-152 0.1 25 0.2 48 0.4 61 0.8 81
57 n-BuCI 0.1 20 0.2 34 0.4 42
58m n-BuC1 0.1 22 0.2 36 0.4 14 0.8 20
'Overall yield from aldehyde. Analytical results are within 0.45% of the theoretical values unless otherwise noted. See experimental Section for protocol mixture of trans/cis 1.4/1. 'Percent inhibition calculated as if contribution of cis isomer was zero Anal caled: C. 60.99. Found C, 68.54. Mixture of trans/cis 28/1. anal Calcd C, 7282 Found: C. 72.27. Mixture of trans/cis 3.8/1. mixture of trans/cis 2/1. *Preparationof aldehyde, mp 112-116 "C;Hennion, G. F.; Fleck, B. R J. Am. Chem. soc. 1955, 77,3263. Preparation of aldehyde, mp 96-98 "C; Bergmann, E, D.; Weiler-Feilchenfeld H.; Mandel, N. Vietnamica Chim. Acto 1966, 129; Chem. Abstr. 1972, 72, 32'764. preparation of aldehyde, mp 42-44 "C;Kohler, E. P.; larsen R G. J. Am. Chem. soc. 1935,57,1452 IIL The decahydronaphthalenes51 and 52 and the ada- binding to HMG-CoA reductase is sensitive (a) to the mantyl compound 53 all possessed similar activity, while stereochemistry of the lactone moiety, bto the ability the cyclohexanes 49 and 50 were less active. Aromatization of the lactone moiety to be opened to a dihydroxy acid. of the cyclohexane ring to give the phenyl derivative 54 (c) to the length of the moiety bridging the lactone and had little effect on activity. However, substitution of the the lipophilic group, and (d) to the size and shape of the bridging moiety with larger aromatic groups such as those lipophilic group. Further modifications of the lipophilic in compounds 55-57 increased activity about 10-fold i.e. groupleading to more potent inhibitors will be described to about 1% of the inhibitory activity of compactin. in subsequent papers from these Laboratories. Scission of the 4a-4b bond of compound 56 provided a less Experimental Section compact molecule 58 with diminished activity. Melting points were determind a Thomas-Hoover The and relative potency values of the most active on capillary IC50 melting point apparatus and are uncorrected. Proton nmr compounds evaluated in this study are compared in Table spectra were recorded in cdciunless noted otherwise onearlier IV. Although the compounds described above are only a varian T-60,em-390 or NT-360spectrometer. chemicalshifts moderately active HMG-coa reductase inhibitors, analysis are reported in parts per million relative to Me,Si as the internal of their intrinsic inhibitory activities suggests that inhibitor standard Elemental analysis for carbon hydrogen, and nitrogen 352 Journal of Medicinal chemistry 1985, Vol. 28, No. 3 Stokker et al
Schema I OH
CI
3 5 I
t 6a(-)
10 8 9 ch2coh2co2ch3 H+. OH'. NaBH,, EtOH. C4H5CH3 A. H,, Rh/C. Dibal. CH3OH, PPTS. NH,.
Table IV. In Vitro Inhibitory Potencies against HMG-CoA (0.125 g) in CH3OH (2 mL) was added dropwise to e stirred Reductase suspension of 2,4-dichlorobenzaldehyde (7.5 g. 0.043 mol) in ac- etaldehyde mL) in an ice bath The resulting IC50 rel (30 cooled solution was stirred 30 min with cooling. diluted with acetic anhydride no. potency µm (25 ml and heated at 120 °cfor 1 b This mixture was cooled, compactin 0.01 100 diluted with H2O (60mL) and 6 N HCI (25 mL) and heated at 6a 22 0.08 100 "C for 0.5 h The light brown, oily product solidified upon 6a(+) 10.8 0.16 cooling It was collected, dried, and triturated with et2o to provide 7 15.2 2 (85 g, mp 106-108 "C. An analytical sample was prepared 13a 20 0.09 98%). by recrystallization hexane to provide 2 as a pale yellow solid: 16 19.8 0.08 from mp 107-108 °c 6.70 dd, J 15.6 (3 47 129 NMR (H, Hz 7.20-7.73 m), 787 (H, d. J 15 d. J anal. 51 107 H. Hz), 9.8 (H, 6 hz 52 90 c9h6cl2o)C,H. 55 1.9 1.1 Methyl (E-7-(2,4-dichlorophenyl)-5-hydroxy-3-oxo-6- 56 0.89 1.5 heptenoate (3) was prepared by a modification of the procedure 57 1.6 0.9 of Weiler.16 Methyl acetoecatate (23.2 g, 0.2 mol) was added dropwise to a stirred suspension of sodium hydride (50% oil relatives precision is the micromolar concen- 10% °i50c is suspension) (10.5 g, 0.22 mol) in anhydrous THF mL) at 0 tration of the inhibitor required to give 50% inhibition under the (500 conditions of the assay system. °cunder a N2 atmosphere. The resulting solution was stirred 15 min at 0 °cand then treated with a 22 M solution (95.4 mL 0.21 mol) of n-butyllithium in hexane over 10 min. The yellow were determined with a Perkin-Elmer Model 240 elemental an- solution was stirred 15 min at 0 °c and then was treated with alyzer and are within 0.4% of theory unless noted otherwise. a solution of 2 (44.2 g. 0.22 mol) in anhydrous THF (260 mL) Optical rotations were determined with a Perkin-Elmer Model The resulting orange solution was stirred 15 min at 0 °cand then 141 polarimeter. all starting materials were commercially available quenched by dropwise addition d12 N HCl 48ml The reaction unless indicated otherwise. mixture was diluted with H2O (300 mL) and extracted with Et2o 3-(2,4-Dichlorophenyl)-2-propenal (2) was prepared by (3 x 300 ml The organic extracts were combined washed with modification of the procedure of Baker.23 A solution of NaOH brine (2 X 200 mL) dried over MgSO,. and filtered. The filtrate was evaporated in vacuo, leaving a red oil. The red oil was stirred in petroleum ether (200 mL) in order to remove the mineral oil. The mixture was cooled and the petroleum ether decanted to ......
Scheme Ill cl on o
11
0 Ac
19
12 14 \ CI 0 20 MeOH,PTSA TiCI,. MeOH. NaBH4 OH-. H+. Scheme IV CHO \ a.b
a Silica gel. t CIS isomer C 13b Scheme 13a V
CHO I
21 22
16 a HO2CCO2H. BrCH,COBr, C5H5N. Zn,CuBr, Et,AICI. Ac2O, C5H5N. I OH". H+. A, C6H5CH >- H,, Rh/C. PTSA, C6H5CH3 A- 24 23 25 (2 H, B). 3.70 (3 H, e), 4.76 H m), 6.13 IH,dd J = 15,6 &), 6.90 (W,d, J = 15 Hz), 7.0-1.5 (3 H, m). hydroxy-2h-pyran-2-one (4). The eater 3 (20g, 6.3 mmol was H cho stirred in 0.1 N NaOH (200 mL) for 4 h. The resulting solution was acidified with 6 N HCI to provide a yellow solid which was recrystallized to analytical purity yield 0.93 g NMR h h 2.60 (2 H, m), 5.00 hs 5.13 (H,m), 4.67 (H, dd, J 15.6 26 27 Hz 6.97 (H,d, J = 15 Hz), 7.27-7.87 (3 H, m), 11.5 (H.br s Methyl a H,, Ru/c EtOH. SOci2 Pd/C, 2,6-(Me)2C5H3N. AICI3 OH-. g H'. heptenoate 5 sodium borohydride (1.3 g. 33.7 mmol) was H,, added with stirring to a cooled solution (5 °c of 3 (10.7 g. 33.7 mmol in EtOH 100 mL) at a rate sufficient to maintain the with (49:1,v/v. 500 mL) provided a forerun which internal temperature at 15-20 °c The resulting solution was was discarded. continued elution with the same eluant (3500 mL) stirred an additional 2 h with ice-bath cooling and then acidified provided 5 as a yellow oil: NMR a 1.60-1.93 (2 H, m), 2.50 (2 H, with 6 N HCL the resultingmixture was diluted with H2O (250 d. J 6 Hz), 3.67 (3 H, a), 4.13-4.77 (2 H,m), 5.93-6.40 (H.m), mL) and extracted with et2o (3 X 200 mL). The et2o extracts 6.93 (H, d, J= 15 Hz) 7.17-7.50 (3 H,m). Anal werecombined, washed with brine,dried overmgso and filtered H; C: calcd 52.68 found, 52.25. The filtrate was evaporated in vacuo to provide a yellow oil (10.4 g, 97%). A portion of the oil was purified by medium-pressure 6aand 6bAn etoh so- chromatography on a 25 x 1000 mm silical gel column. Elution lution (200 mL) containing 5 (8.4 g, 26.3 mmol) and 1 N NaOH 354 Journal of Medicinal chemistry 1985 Vol. 28, No. 3 Stokker et al. Scheme VI organic extracts were washed with brine. dried over MgSO4 and filtered the filtrate was evaporated in vacuo to provide a mixture of acid and lactone (7.8 g, 97%). A solution of this mixture in J" toluene (100 mL) was heated at reflux in a Dean-Stark apparatus. After 2 h, the Dean-Stark apparatus was replaced with a Soxhlet 3-A molecular sieves (100 g The Solution was refluxed for an additional 4 h and then the toluene was removed in vacuo leaving a yellow oil (7.2 g, 95%) which was a mixture of 6a and The oil was chromatographed a silica gel column 28 29 6b on (500 g Elution with CH2cl2-acetone (4:1, v/v 900 mL) provided a forerun which was discarded. Continued elution with the same eluant (300 mL) gave the trans isomer 6a (2.5 g Recrys- tallization of the solid provided an analytical sample, as colorless CN cho needless NMR (acetone-d6 6 2.06 (2 H. m), 2.69 (2 H, m), 4.43 (H.m). 5.42 (H,m), 6.49 (H,dd, J 15,6 Hz), 7.08 (H,d, J 15 Hz), 7.33-7.59 (2 H. m), 7.79 (H,d, J 8 Hz). An isomeric purity of 99.8% was determined for 6a by HPLC on a 31 32 33 Whatman Partiail-5 RAC column with 15% 2-propanol/hexane as the eluant. The time of elution was 4.96 min at a flow rate of 6 mL/min. further elution of the column with the same eluant (600 mL) gave the cis isomer 6bas a solid (1.25 g). Recrystallization gave an analytical sample as colorless needless nmr (acetone-d6 34 36 1.50+93 (4 H, m), 4.36 (H,m), 5.02 (H,m), 6.37 (H, dd, J = Dibal. 15,6 Hz). 7.02 (H, d, J 15 Hz),7.16-7.50 (2 H, m), 7.67 (H,d, NaCN. J 8 Hz). An isomeric purity of 993%was determined for 6b by HPLc on a Whatman Partisit-5 RAC column with 15% 2- Scheme vii propanol/hexane as the eluant. The time of elution was 5.79 min at a flow rate of 6 mL/min cl cl 4-hydroxy-2H-pyran-2-one (7). A solution of 6a (1.5 g, 5.2 mmol) io THF (100 mL) was stirred magnetically and hydro- genated at room temperature under atmospheric pressure in the presence of5% rhodium on carbon (150 mg) until 1.25 molar equiv 36 37 of hydrogen had been consumed. After removal of the catalyst brCH2CH(OEt) 2N H2SO4 by filtration, the filtrate was evaporated in vacuo, leaving a solid. The solid was recrystallized to provide 7 (0.9 g nmr 6 1.67-2.17 Scheme viii (4 H, m), 260-3.13 (4 H,m), 4.30-4.50 (H,m), 4.57-4.90 (H, m), 7.14-7.44 (3 H, m). CI Resolution of (6a), 2 cho A solution of 6a (2.87 g, 10 mmol in (R)-(+)-a-methyl- benzylamine (15 mL) stirred for 18 h at ambient temperature CI was and then poured into H2o (100 mL). This aqueous mixture was 38 acidified with 6 N HCI and extracted with et2o (3 X 100 mL). CI The et2o extracts were combined, with brine, dried over I Cl washed MgSO4 and filtered. Evaporation of the filtrate in vacuo provided the crude diastereomeric amides as a tan viscous oil (4.1 g, 100%). this oil 3.1 g, 7.6 mmol was chromatographed on a silica gel 200 g). Elution With aceton-cH,Cl2 1:4, v/v 1200 mL) 39 40 gave a forerun which was discarded Continued elution with the same eluant provided the mixture of diastereomeric amides as a viscous oil (3.0 g, 97%). This mixture was separated by chro- matography on a Waters Prep lc 500. The separation was accomplished by using two prep PAK-500 silica cartidges in series and eluting with acetoneCH2Cl2 (1:4, v/v). Use of the shave- recycle technique provided diastereomer A (1.36 g) and diaste- reomer B (1.2 g). recrystallization of diastereomer A from n-butyl chloride gave colorless clusters (1.0g) which melted at 106-108 °c NMR 6 1.47 42 (3 H, d, J 6 Hz). 1.70 (2 H,m), 2.33 (2 H,d, J 6 Hz). 4.30 CI (H,m), 4.58 (H.m). 5.13 (H, m), 6.20 (H,dd, J 15,6 Hz). 6.33 I (H, m). 6.93 (H, d, J 15 Hz), 7.33 (8 H. m). Anal. CI2No5) C. H. N. recrystallization of diastereomer B from n-butyl chloride gave a pale yellow solid which melted at 55-60 °cNMR 6 1.47 (3 H, d. J 6 Hz), 1.70 (2 H,m), 2.33 (2 H,d. J 6 Hz). 4.33 (H.m), 4.60(H,m),5.16(H,m).6.17(H,dd.J=15,6Hz),6.23(H,m), 6.93 (H.d J 15 Hz).7.33 (8 H, m). Anal. C, H, N. trans E)-6-[2-(2.4-Dichlorophenyl)et hen (6a(+)). Diastereomer A (0.74 g, 1.8 mmol) was dissolved in 95% ethanol (25 mL) containing n ... h mg coa Reductase inhibito
The resulting mixture was extracted with Et2o The Et2o extracts H. d. J 6 Hz),4.73 (H,m), 6.10 (H, dd). Anal. were combined. washed with brine, dried over MgSO4 and filtered C, H. The filtrate was evaporated. leaving the intermediate acid as a 6-(2,4-Dichlorophenyl)-4-hydroxy-5-hexen-2-one 2-bromoacetate yellow oil (0.54 g). A solution of the oil in toluene (150 mL) was (12). 2-Bromoecetyl bromide (1.1 mL, 13.2 mmol) was added refluxed through a soxhlet containing molecular sieves (3 A) for dropwise to a stirred solution of 11 (3.4 g, 13.1 mmol and pydine 5 h. The solution was evaporated in vacuo, leaving the lactone (1.07 ml 13.2 mmol in Et2o (100 mL) at 0 "C. The ice bath 6a as a yellow solid Recrystallization gave colorless needles was removed and the reaction mixture was stirred at 20 "C for (0.13 g [a]25D +5.9° (c0.425,CHCl3 NMR 6 203 (2 hm). 2.73 2 h and then diluted with H2o (100 mL). The organic layer was (2 H, m), 4.46 (H. m). 5.41 (H,m), 6.19 (H, dd, J = 15,6 Hz),7.01 separated and washed with 1 N HCI (100 mL), H2o (2 x 100 mL) (H, d, J 15 Hz), 7.14-7.50 (3 H. m). and saturated brine, dried (MgSO4 filtered, and evaporated. the (-)-trans-(E)-6-[2-(2.4-Dichlorophenyl)et residual oil was chromatographed on silica gel. Elution with rahydro-4 (6a(-)). Disstereomer CH2cl2-acetone 99:1v/v; 1.9 L) provided 12 (2.8 g, 56%): nmr B (1.1 g, 27 mmol was dissolved in 1 N NaOh (5.4mL, 5.4 mmol 6 2.2 (3 H, I), 2.92 (2 H, t, J 7 Hz),3.85 (2 H, s), 5.9 (H, m and the solution was refluxed for 18 h. The same workup and 6.15 (N.m), 6.95-7.5 (4 H,m). Replacement of pyridine with an lactonization used in the synthesis of 6agave 6a as a yellow equivalent of TEA provided only the dienone (6-(2,4-dichloro- solid. Recrystallization provided colorless needles (0.34 g mp as a yellow powder (77%mp 80-82 114-115 "C;[a]25D -6.6° (e 0.555, CHCl3 NMR 6 2.03 (2 H,m), "C; NMR 6 2.28 (3 H,s 6.15 (H, d, J 15 Hz),6.7-7.6 (6 H. 2.73 (2 H,m). 4.46 (H,m), 5.41 (H.m), 6.19 (H, dd, J 15,6 Hz), m). 7.01 (H,d, J 15 Hz), 7.14-7.50 (3 H, m). The optical purities of 6a and 6a(-) were determined by NMR with ca. 0.5 molar (13). A solution of 12 (2.8 equiv of Eu(hfbc)3 in cdcl3 each enantiomer was found to be g, 7.4 mmol in dry THF (50 mL) was added dropwise to a vig free of the other enantiomer within the limits of detection orously stirred slurry of activated Zn dust (720 mg. 11.1 mmol). (threshold ca 2%). Therefore. the optical purity of 6a and CuBr (60 mg, 0.4 mmol), Et2a1cl (25% solution in toluene; 3.2 6a(-) was estimated to be 98 2%- mL8 mmol), and dry THF (50 mL) under N2at 20 'C. Stirring (2R and (2S*,4R*,6S was continued for 5 h before quenching with pyridine (8 mL) c followed by H2o (500 ml addition and Et2o extraction (3 x 150 hydropyran (8 and 9). A solution of Dibal (0.89 M)in hexane mL). The combined Et2o extracts were washed with 1 N HCI (4.31 mL, 3.83 mmol was added dropwise over 10 min to a stirred (2 x 50 mL), H2o (2 X 250 mL), and saturated brine, dried toluene (20 mL) and THF (10 mL) solution of 6a (0.5 g, 1.74 (MgSO4 filtered. and evaporated, leaving a sticky, pale yellow mmol) which was cooled to -78 'C. After stirring another 30 min solid (1.8 g) which was a mixture of 13a and 13b. This crude at -78 °cthe reaction was quenched by the addition of MeOH product was digested once with Et2o (40 mL) and then (613 mg, 19.15 mmol). After the reaction mixture had warmed from n-butyl chloride 25 mL) to provide the trans isomer 13a to ambient temperature, it was diluted with H2o (1 mL), treated as tiny colorless crystals (550 mg): mp 136-138 °cNMR 6 1.4 with Celite, and stirred for an additional 15 min. Anhydrous (3 H. s ch3 1.73 (5-H,dd, J 11, 13 Hz), 2.06 (5-H,ddd, J Na2-so4 was added and stirring was continued another 15 min. 2, 11, 13 Hz), 2.12 hs OH),2.45 (3-Hd, J 18 Hz),273 This mixture was filtered and the filtrate evaporated in vacuo (3-H dd, J 2 18 Hz),5.2-5.45 (6-H, m), 6.13 h dd, J 15, to provide a mixture of lactols (0.5 g, 99%). A CH3OH (10 mL) 6 Hz),6.92 (H, d, J = 15 Hz),7.0-7.43 (3 H, m). solution of the lactol mixture (0.25 g. 0.86 mmol was treated with thefiltrates from digestion and crystallization vide supra were PPTS (25 mg) and then refluxed through a Soxhlet containing combined. evaporated, and chromatographed with a Waters prep molecular sieves (3 A). The reaction was cooled and diluted with LC 500. The separation was accomplished by using two prep Et2o 200 mL). After washing with saturated NaHCO3 solution, PAK-500 silica cartridges in series and eluting with CH2Cl2 H2o and brine, the organic layer was dried over MgSO4 and acetone (15:1,v/v). By use of the shave-recycle technique, the filtered. The filtrate was evaporated in vacuo to provide the cis (b. 220 mg) and the trans (a, 230 mg) isomers of 13 were mixture of 8 and 9 255 mg, 78%). separated The cis isomer (13b) was crystallized from n-butyl This mixture was separated byflash chromatography an a silica chloride-hexane (2:1,v/v) (120mg): mp 135-137 °cNMR 6 1.46 column. Elution with CH2cl2-acetone (19:1v/v) first provided (3 H, a, CH3),1.8-24 (3 H. m,5-H, OH), 2.65 2H, 3-h the minor isomer 9 followed by the major isomer 8. 4.8-5.1 (6-H,m), 6.26 (H, dd, J 15,6 Hz), 7.03 (H, d, J 15 Isomer 9: NMR 6 1.47-2.13 (4 H. m). 3.44 (3 H. s 3.53 (H, Hz). m),4.13 (H, m), 4.77 (W,m), 6.20 (H,dd. J 15,6 Hz),6.98 (H. The epimeric alcohols were readily distinguished by analytical d, J = 15 Hz), 7.13-7.57 (3 H, m). TLC (fluorescent silica gel (40 A), 1 x 3 in., MK6F, Whatman isomer 8: NMR 6 1.47-1.93 (4 H,m), 3.53 (3 H,s 4.40 (H, with CH2cl2-acetone (9:1, v/v) as the eluant; R, 0.25 for 13b and m), 4.63 (H, m), 4.85 (H. dd, J 9.3 Hz). 6.23 (H, dd, J = 15, 0.30 for 13a. 6 Hz),6.98 (H,d, J 15 Hz),7.10-7.57 (3 H. m). Alternate Route to 13. 4-Acetoxy-6-(2,4-dichloro- erytlro E)-7-(2.4-Dichloropheny roxy-6- (14). Acetyl chloride (12 ml 165 mmol tenamide (10). Lactone 6a (6.0 g. 20.9 mmol) was dissolved was added dropwise to a stirred solution of 11 (3.9 g. 15 mmol) in dry MeON (100 mL) and saturated with anhydrous NH3 at in pyridine (60 mL) cooled at 0 'C. The ice bath was removed 0 'C. The reaction was sealed and stirred at Mom temperature and the reaction mixture was stirred at 20 "C for 2 h and then for 13 h. stored in the freezer overnight. and then worked up by diluted with Et2o (300 ml The Et2o solution was washed with evaporation under reduced pressure Two recrystallizationsrecrystallizations from 1 N HCl(3 X 300 mL) and saturated NaHCO3 dried (MgSO4 acetone-hexane gave 10 (4.0 g): mp 117-118 °cNMR 6 1.72 (2 filtered and evaporated The residual pale amber oil (4.1 g) was H, m). 2.40 (2 H. d. J=6 Hz). 4.22 (1 H, m),4.55 (1 H, m), 5.00 chromatographed on silica geL Elution with CH2cl2 (2L) provided (2 H. t, J 5 Hz). 6.17 (1 H. d, J 7 Hz),6.35 (1 H, d, J 7 14 as a pale yellow oil (3.95 g, 87%); NMR 6 2.03 (3 H, s217 Hz), 6.8-7.75 (3 H, m). (3 H, s 283 (2 W, dd, J 7, 2 Hz). And. C, H 6-(2,4-Dichlorophenyl)-4-hydroxy-5-hexen-2-one (1 1). Ethyl 2-acetorypropene (3.3 ml 30 mmol and tri-n-butyltin methoxide methyl-6-heptenoate (15). A solution of 14 (1.3 g, 4.3 mmol (5.7 g, 24 mmol) were combined and stirred at 60-70 "C under and ethyl bromoacetate (0.47 mL, 4.2 mmol in dry THF (10 mL) N2for 1 h and then placed under vacuum for an additional 30 was added dropwise to a vigorously stirred slurry of activated Zn min. Propenal 2 (4 g, 20 mmol was added and the reaction dust (490 mg, 7.5 mmol CuBr (29 mg, 0.2 mmol), Et2alcl (25% mixture was stirred at 70 °c under N2 for 4 h. The clear reaction solution in toluene 1.72 mL, 4.3 mmol and dry THF (5 mL) mixture was then oooled, treated with malonic acid (1 g, 10 mmol under N2 at 20 °c Stirring was continued for 5 h before in Et2o (20 mL), and refluxed for 30 min. After cooling to -20 quenching with pyridine (3.5 mL) followed by H2o (50 mL) 'C, the reaction mixture was filtered and the precipitate was addition and Et2o extraction (3 X 80 mL). The combined et2o washed with (4 x 10 mL,). The Et2o solutions were mm- extracts were washed with 1 N hCl(2 X 25 mL), h2o (2 X 50 bined and evaporated, and the residual oil was chromatographed mL), and saturated brine dried (MgSO4 filtered and evaporated on silica gel Elution with CHcl3-M3OH 99:1v/v. 2 L) provided leaving crude 15 as a pale yellow oil (1.2 g, 75%); NMR 1.28 I I sd a thick yellow oil (4.2 g, 81 %); NMR 6 2.2 (3 H. s 273 (2 (3 H, t, J 7 Hz), 1.33 (3 H,s 2.10 (3 H, a), 2.45 (2 H, d J 356 Journal of Medicinal Chemistry, 1985. Vol. 28, No. 3 Stokker et al. 13 Hz), 4.16 (2 H,q, J 7 h2 mg under hydrogen in dry THF (80 mL) containing 2,6-di- Compound 13. This compound was prepared similarly to 6. methylpyridine (2.14 g, 20 mmol). Hydrogen uptake occurred With 15 (1.2 g, 3.2 mmol) as starting material. 13 (120 mg) was at a steady rate and was complete in 1 h. The reaction mixture obtained as a mixture of trans (54%) and cis (46%) isomers as was filtered and the filtrate was evaporated. The residue was determined by hplc (Whatman Partisil-10 PAC eluting with distributed between 5% HCI (200mL) and et2o (200 mL). The i-PrOH-hexane (1:10,v/v) at 2.1 ml/min). Times of elution are et2o wan washed with aqueous NaHCO3, H2O and saturated 3.0 min for a and 4.0 min for b. brine, dried (MgSO4 filtered, and evaporated. The residue was distilled in vacuo to provide 25 as a clear colorless oil (27g, 69.5%): (16). This compound was bp 96-99 "C (0.5 mm); NMR 6 1.0-2.0 (19 H, m 2.50 (2 H,dt, prepared analogously to 7, starting with 13a (100 mg,033 mmol J 8,1h2 9.8 (N,t J = 1 Hz). Anal. (C13H22O) H;C: celcd The residual oil was chromatographed on silica gel Elution with 80.35; found, 79.70. CH2C12-acetone (19:1,v/v; 350 mL) provided 16 (50mg, 50%): 3-( trans-syn Acid NMR 6 1.37 (3 H,s 1.621 (4 H,m), 25-3.1 (4 H, m), 4.55-4.90 (26). Ethyl propanoate (24) (24 g, 100 mmol) was mixed with (H, m), 7.15-7.25 (2 H, m). 7.33-7.43 (H,m). anhydrous AICI3 (27 g, 200 mmol at 0 "C and then stirred at 20 (E) 6- 2-(2.4 hlorophenyl)e t hen 4 "C for 70 h The viscous reaction mixture was cooled to 0 "C, methyl-2R-pyran-2-one (17). A solution of 13a (150 mg, 0.5 quenched with ice-cold 1 N HCl (500 mL), and extracted with mmol) in toluene (75 mL) containing PTSA (5 mg) was refluxed et2o (4 X 100 mL). the et2o extracts were combined and through a Soxhlet filled with 3-A molecular sieves for 6 h. After evaporated to afford a residue which was stirred with 1N NaOH evaporation of the toluene, the residue was chromatographed on 100 mL, 100 mmol) and MeOH (400 mL) for 2 h at 20 "C. The silica geL Elution with CHC13-Meoh (991. v/v. 60 mL) provided reaction mixture was evaporated and the residue was distributed 17 (130 mg, 90%): mp 108-110 "C; NMR 6 2.0 (3 H, s2.4-2.6 between H2O (200 mL) and et2o (200 mL). The aqueous layer (2 H, m), 5,0-5.25 (H,m), 5.85-6.0 (H,m), 6.25 (H,dd, J 16, was cooled to 0 "C and acidified with 12 N HCl to provide 26 as 6 Hz). 6.95-7.6 (4 H, m). a sticky yellow solid. Recrystallization from petroleum ether gave Dimethyl Acetal (18). A 9.1 g (43%), mp 98-98.5 'C. Anal. (C13H22o2)C, H. solution of 2 (10 g, 50 mmol ptsa (50 mg), and anhydrous 3-( trans (27). MeOH (100 mL) was refluxed through a Soxhlet filled with 3-a This compound was prepared similarly to 25,starting with 26 (10.1 molecdar sieves for 1 h. The solution was cooled, treated with g, 48 mmol). The intermediate acid chloride was obtained as a anhydrous K2CO3 filtered and evaporated The residual oil was clear colorless liquid (10.2 g, 93%): bp 113-116 °c (0.8-1.0 mm); chromatographed on silica gel Elution with CH2cl2 provided 18 NMR 6 0.6-1.9 (19 H, m), 2.88 (2 H, t, J 9 Hz). Anal. (CIJ- as a clear, pale brown oil (10.4 g, 84%). H21CIO) C. H. Reduction provided 27 (74%): bp 107-112 "C Methyl (1.3-1.5 mm); NMR 6 0.5-1.8 (19 H. m), 243 (2 H,dt, J 8,1 noate (19). A solution ofticl in CH2CI2 (3 M, 10 ml30 mmol Hz),9.85 (H,t, J 1 Hz). And. (C13h22O)H; C: calcd 80.35: was added dropwise to a Vigorously stirred solution of 18 (6.18 found. 79.94. g, 25 mmol) in CH2C12 (40 mL) at -78 "C under N2 A solution (E)-3-( (28). A mixture of of diketene in CH2cl2 (5 M. 10 mL. 50 mmol) was added in a (156 g. 1.0 mol), cyanoacetic add (78 g, 0.9 mol). steady stream. The dark reaction mixture was stirred at -78 °c and ammonium acetate (3 g) in toluene (200 mL) and pyridine for 1 h and then anhydrous MeOH (20 mL) was added. After (100 mL) was heated at reflux for 2 days; water was collected in stirring for an additional 30 min at 20 "C, the light yellow solution a dean-stark trap. After evaporation of the solvents, the product was poured into cold, aqueous K2CO3 (10 g/L). The yellow salts was obtained by distillation in vacuo (~140-185 "C, 0.7 mm were separated by filtration and washed with CH2cl2 The organic Recrytallization from EtOH-H2O gave 28 (145.7 g, 88%): mp layers were combined, washed with H2O and saturated brine, dried 69-71 'C; NMR 5.94 (1 H, d, CH, J 17 Hz), 7.25-8.1 (7 H, (MgS04 and evaporated. leaving 19 as an orange oil (6.6 g, 80%) m, C10h7 8.25 (1 H, d, CH, J 17 Hz). And (C13H9n) C, H, which exhibited one major spot (RtO.45) on TLC (fluorescent silica N. gel (40 A), 1 x 3 in. MK6F, Whatman) after eluting with 3-( (29). A mixture of 28 CH2Cl2-MeOH 99:1 v/v): NMR 2.90 (2 H, d, J 4 Hz), 3.43 (17.9 g. 0.1 mol) and 5% Pd/C (1.0 g) in EtOH (200 mL) was (3 H. s 3.63 (2 H,s 3.83 (3 H. s 4.13-4.57 (H, m), 6.10 (H, dd, hydrogenated in a Parr apparatus. After fitration and removal J = 15,7 Hz), 6.83-7.43 (4 H, m). of the solvent, the product was obtained by distillation in vacuo Acid (23) 150-165 "C, 0.5 mm). Recrytallization from et2o-petroleum and Ethyl Ester 24. acid (79.6 g, ether afforded 29 (16.4 g, 90%): mp 48-51 'C; NMR 6 2.72 (2 400 mmol) was hydrogenated EtOH over 5% Ru in (600 mL) on H, t, J 8 Hz),3.4 (2 H,t, J 8 Hz), 7.25-8.1 (7 H, m). Anal carbon (8.0 g) at 100 °c and 3000 psi The catalyst was removed (C13h22N)C. h N. by filtration and the filtrate was evaporated. The residue was (30). To a stirred suspension distributed between (700ml and NaOH (2 X 400 mL). et2o 6% of 29 (21.7 g. 0.12 mol) in et2o (400 mL) at -78 "C under N2 was The et2o layer was washed with H2O and saturated brine, dried added a solution of Dibal(85 0.128 mol) in toluene (MgSO4 filtered, and evaporated. Distillation of the residue ml (25.3%. 0.8466 g/mL) over a period of 1 h After an additional 1 h at -78 provided ethyl as a dear colorless liquid g. 50.7%): ester 24 (48.4 "C, the dry ise-acetone bath was removed and the reaction bp 1.0-1.9 2.27 (2 H, 120-122 "C (1.0 mm): NMR 22 H,m). mixture was stirred at room temperature for 3 h. The mixture t, J 6 H, q, J And. (C15H26o2) C, H. Hz), 4.14 (2 6 Hz). was added slowly to 5% aqueous H2so4 and then extracted with The basic layer vide supra was extracted with et2o (200 mL) The solution was dried and evaporated and the and then acidified with N HCl after cooling to 0 "C. The acid et2o et2o 12 residual distilled to give the product a colorless oil. bp was removed by filtration and petroleum ether oil as crystallized from 113-117 °c (0.2 mm), which solidified on standing. Recrytallization to provide 23 as colorless g, mp 80-82 needles (23.2 27.6%): "C; lization from et2o-petroleum ether yielded 30 g, 72%): mp 6 0.8-1.8 W, 2.33 (2 t, J 8 11.4 br (15.8 NMR (19 m), H, h2 (W, s 29-31 °c 2.82 (2 3.4 (2 J 8 AnaL (C13H22o2) NMR H, m), H,t h2 7.21-8.02 C, H. 9.86 (1 t, j 1 Anal. (C13H12o)C, H. (25). (7 H,m), H, Hz). A mixture of 3126 Propanoic acid (23) (10g, 47.6 mmol) was dissolved in SOC12 (20 4 (32). (58.8 g, mol) and sodium cyanide (25 g, 0.5 mol) in EtOH mL) and the solution was heated at reflux for 3.5 A The solution 0.24 was evaporated and the residual oil was distilled in vacuo to (300 mL) and H2O (100 mL) was heated at reflux with stirring for 5 The reaction mixture was concentrated in vacuo provide the acid chloride as a clear colorless oil (7 g. 64%): bp h cooled, and extracted with solution was dried, filtered, 113-120 °c (0.8-1.0 mm); 1.0-2.0 (19 m). 2.87 (2 H, et2o The et2o NMR H, and concentrated to give the crude product, which was purified t, J = 8 Hz), Anal. (CI3HmCIO)C. H. g The acid chloride (4.56 g, 20 mmol) was added dropwise to a by distillation: yield 38.8 (84%); bp 151-155 "C (0.3 mm well-stirred suspension of hydrogen-equilibrated 10% Pd/C (200 . HMG-coa reductose lnhibitors Journal of medicinal chemistry 1985, Vol. 28 No 3 357
1.7-2.3 (4 H, m), 3.08 (2 H, t,J = 7 Hz),7.1-8.0 (7 h m). Anal. and purified through the second ammonium sulfate precipitation (C14H13H) C. H. n step as described by Kleinsek et al." The enzyme preparation 4-( this product was prepared was tested for HMG-coA reductase potency and diluted with 100 from 32 in a manner similar to that used for 30 and purified by mM phosphate buffer (pH 7.2) so that 100 µl of the enzyme distillation: bp 123-126 °c (0.1 mm); NMR 6 2.13 (2 H,m 2.50 solution, when added to the assay control, gave a value of (2 H,m), 3.12 (2 H,t, J 8 Hz). 7.2-8.3 (7 H, m), 9.82 (1 H, t, 50000-60000 dpm. The enzyme preparation was stored at -80 J = 1 Hz). "C. wan prepard analogously to HMC-CoA Reductase Inhibition Assay. The assay is es- 33 starting with 3427 (4.73 g, 25 mmol Distillation under vacuum sentially the procedure of Shefer et al.35 The complete assay provided 35 as a colorless oil (27 g, 56% bp 107-115 °c(1.0-1.6 medium contained the following in a total volume of 0.8 mL: mm) lit.bp 101-103 "C (1.5 mm)];NMR 1.3-2.1 (17 H, m), phosphate buffer, pH 7.2.100 mM,Mgcl2 3 mM;NADP, 3 mM; 2.4 (2 H. dt, J = 9,0.8 Hz), 9.85 (H.d. J 0.8 Hz). glucose6-phosphate, 10 mM, glucose-6-phosphate dehydrogenase, (37) was prepared by 3 enzyme units; reduced glutathione, 50 mM; HMG-CoA glu- the method of Julia and Tchernoff29 New England Nuclear), 0.2 mM (0.1 µci and partially (a) diethyl acetal: purified enzyme stock solution, 100 µl yield (45%); bp 140-142 'C (22 mm) lit.29 bp 137-140 'C (2 Test compounds or compactin (after first being converted to mm)];NMR 1.23 (6 H,t, J 6 Hz), 3.43-3.93 (4 H, m), 4.0 (2 the sodium salt of their dihydroxy acid form in situ by addition H, d. J 4 Hz), 4.80 (H.t. J 4 Hz),6.67-7.3 (3 H. m of 1 N NaOH (1 equiv)) were added to the system in 10-µl (b) 37 yield (80%);bp 125-128 °c(2.2 mm) lit.29 bp 120 °c volumm at multiconcentration levels, After a 40-min incubation (1.5 mm NMR 6 4.60 (2 H, s6.67-7.4 (3 H, m), 9.8 (H,s at 37 "C with shaking and exposure toair. the reaction was stopped (41) was synthesized from by the addition of 0.4 mL of 8 N hCI. after an additional 30-min 2 by the procedure used by Men and Edens22 to convert 3- incubation period at 37 °cto ensure the complete lactonization phenyl-2-propenal to of mevalonic acid to mevalonolactone,0.2 mL of the mixture was (a) (38): added to an 0.5 x 5.0 cm column containing 100-200-mesh bio- yield (67%). An analytical sample was crystallized from cyclo- Rex 5, chloride form (Bio-Rad), wetted with distilled water, as hexane-toluene: mp 125-126 °cNMR 6 7.35-7.65 (2 H, m). 8.2 described by Alberta et al.9 The unreacted 14C]HMC-CoA was (H,d. J 6 Hz), 8.25 (H,s 9.55 (H, s Anal. (C9h5BrC12o) absorbed on the resin and the [14C]mevalonolactone was eluted c, H. with distilled water (2 x 1 mL) directly into 7-mL scintillation (b) diethyl vials Five milliliters of Aquasol-2 (NewEngland Nuclear) was acetal (39): yield (70%); bp 130-140 °c(0.1-0.05 mm) as a pale added to each vial, and radioactivity was measured in a Packard yellow oil; NMR 6 1.25 (6 H, t, J 6 Hz). 3.65 (4 hdq, J 6, Tri Carb prias scintillation counter IC50 values were determined 3 Hz),5.0 (H, s 7.2-7.75 (4 H, m). by plotting percentage inhibition against test compound con- (c) diethyl acetal (40): centration and fitting a straight line to the resulting data by yield 88%bp 122-124 'C (0.04 mm); NMR 1.23 (6 H, t J the least-squares method, for estimation of relative inhibitory 6 Hz), 3.5-3.95 (6 W, m), 5.5 (H, s), 7.1-7.5 (3 H, m anal potencies, compactin was assigned a value of 100 and the IC50 (C13H14Cl2o2) C. value of the test compound was compared With that of compactin d41: yield (67%). An analytical sample was cryetallized from determined simultaneously. hexane: mp 57-58 6 7.2-7.7 (3 m), 9.5 (H. s). anal "C; NMR H, Acknowledgment. are indebted Drs. c9h4cl2o c, h We to R F. was prepared by Hirschmann and P. S. Anderson for their encouragement the general procedure of Woflenberg30 n-Butyllithium (272 throughout the course of this investigation, to Dr. W. C. mmol) was added dropwise to a solution of cis-l-ethoxy-2-(tri- Randall and staff for analytical support, to Drs. B. H. (2.3 g. 6 mmol in dry THF (20 mL) Arison and D. W. Cochran for NMR spectra, and to M. at -78 "C. After stirring at -78 'C for 1 h, a solution of 4- z Banker for manuscript preparation. g, in phenanthrenecarboraldehyd32 (1.1 5.3 mmol) dry THF (10 NO. 2,70017-27-5; (&)-3,93863-41-3; (4,93863- ml added rapidly mixture was stirred at Registry was and the reaction 42-4; 5.9363-43-5; 86097-37-2 6a78444-38-9; (-)-6a, -78 for additional 2 min the cooling bath 6a "C an before removing 78444-39-0; 86097-38-3 7,86097-55-4; 893863. after stirring at for 30min reaction mixture was 6b 22°C the treatad 44-6, (lactol), 93863-87-7; (lactol), saturated mL) and the reaction mixture was 8 9,93922-58-8; 9 with NaHCO3 (20 93922-60-2 1093863-45-7; 1193863-46-8; 12 distributed between Et2o 100ml (100 mL). The and h2o Et2o 93863-47-9; 13a,93863-48-0 13b 93863-49-1; 14 layer was washed with h2o (3 X 100 mL), dried (MgSO filtered, then evaporated. Chromatography of the biphasic residue 93863-50-4 15,93863-51-5;16,93863-526; 17,93863-53-7 and 18,93863-54-8; (5)-19,93863-55-9 93863-56-0 on silica gel with elution by ch2cl2 provided 55a (900 mg,73%): threo-20 93863-57-1; 21, 13026-12-5; 23 mp 112-120 °c crystallization from hexane yielded analytically 66-77-3;22, 93863-58-2; (add chloride), 93863-59-3: pure 55a mp 125-129 6.8 (H, dd, = 18.8 23 24,93863-60-6 °cNMR J Hz). 25 93863-61-7; 26 93863-62-8; 26 (acid chloride), 7.5-8.45 (10 H, m), 9.9 (H, d, J 8 Hz). anal (C17H12O) C, H. isolation of HMG-CoA reductase Male Holtzman 93w-63-9 (&)-27,93863-64-0;28, 93863-65-1 29,70067-70-8; 30, 53531-16-1;31, 27650-86-8; 32, 93863-66-2; 33, 93863-67-3; 34, Sprague-Dawley rata (225-250 g were kept on reversed lighting 35 18228-55-236,120-83-2 31,17944-27-3;37 and fed Purina rat chow containing 3% cholestyramine for 7 days 52582-89-5; (diethyl preceding their sacrifice co2 asphyxiation. Livers were acetal 78830-79-2;38,93863-68-1; 39,93863-69-5 40,93863-70.8; by re- 41, 93863-71-9; 93863-73-1; DL-44, moved 6 h into the dark cycle immediately prepare 4293863-72-0 43 and used to 86097-50-9; 45 86097-49-6, 46 93863-74-2; 47 microsomes HMG-CoA reductase solubilized from the freshly prepared the method of Heller and Shrewsbury,33 93863-75-3; 4893863-76-4 4993863-77-5; 50 microsome by 93922-59-9; 5193863-78-6; 52 93863-79-7; 53 93863-80-0; 54 86118-13-0 55 93863-81-1; 55a 27 McKervey, hi. a J. chem Soc., Perkin trans 1972,2707. 93863-82-2 56 93863-83-3; 57 93863-84-4 58 (28) Stepanov, F. N.; dovgan N. L Zh. Org. Khim. 1968,4, 277 93863-85-5; BrCH2CH(OEt)2 2032-351: ch3cho 75-07-0; Chem. Abstr. 1968,6B, 104612w. CH3COCH2cO2H3 105-45-3; ch2=c(oac)ch3 108-22-5 (29) Julia M.;Tchernoff G. bull Soc. chim R.1953.479; chem CH2(CO2H)2 141-82-2; BrCH2cOBr, 598-21-0; BrcH2CO2et Abstr. 1954,48,3930g (30) Wollenberg, r hAlbizati k F; peries R J, Am. Chem. soc 1977,99,7365. (33) heller R A.; Shrewsbury, M. A. J. Biol. Chem. 1976, 251, (31) Leusink. A. J.; Budding, H. aMaraman, J. W. J. Organomet 3815 chem 1967,9,285. Leusink. a J.; Budding, H. A; Drenth. (34) Kleinsek D. A.; Ranganathan S.; Porter, J. W. Proc, natl. W. Ibid. 1968, If, 541 Acad. sci U.S.A. 1977, 74, 1431. (32) Horgan, S. W; morgan D. D; orchin M.J. Org. Chem. 1973, (35) Shefer, S.; Hauser, S.; Lapar, V; Moebach, E H. J. Lipid Res. 38,3801. 1972, 13, 402. 358 J. med. Chem. 1985,28, 358-362
105-362;NCCH2Co2h 372-09-8; dihydro1y-7-(2&dichlorophenyl)-a(E)-hept.enamid~,93922-57-7; 64724-29-4; 2,4-dichlorobenzaldehyde, 874-42-0; 93863-86-6: diketene. 674-82-8; 4-phenanthrenecarboxaldehyde, 41498-43-5 HMG-CoA 93922-56-6; reductase, 902835-7.
Acyclic Analogues of 2’-Deoxynucleosides Related to 9 as Potential Antiviral Agents1,2
John C. martin Gary A. Jeffrey, Danny P. C. McGee Michael A. Tippie, Donald F. Smee, Thomas R. Matthews, and Julien P. H. Verheyden Syntex Research, Palo Alto, California 94304. Received June II 1984
A series of acyclic analogues of related in structure to dhpg 1) have been synthesized and evaluated for antiviral activity against herpes simpler virus type 1 f strain Additionally, the ability of these analogues to function as substrates for the vim-specified thymidine kinase was examined. Phosphorylationby this kinase is essential for antiviral activity. Although the acyclic 4-oxopyrimidine nucleosides were substartes for the kinase, they were devoid of antiviral activity. In the purine series, most analogues similar in structure to DHPG did exhibit significantly lower antiviral activity, indicating that even small modifications in the purine substituents subtantially reduce the antiviral potency. The most active agent, 2,6-daminopurine 27 was only poorly phosphorylatedby the viral kinase; therefore, its activity was most likely due to a prior enzymatic deamination to give DHPG Evaluation of 27 in a mouse encephalitis model has shown it to be nearly as potent as DHPG (1).
The synthesis of Scheme I guanine dhpg 1a potent antiherpetic agent, was re- cently reported independently by us‘ and others DHPG
,
1 is a member of a class of selective antiherpetic nucleoside analogues6 which includes methyllguanine (acyclovir),’ deoxyuridine (BVDU)8 and Scheme II (FIAC)9 Of this class, 0 s DHPG appears exceptionally promising, being found to be effective against not only herpes simplex Virus types 1 and 210 but also cytomegalovirus,10,11a varicella-zoster,11b and Epstein-Barr virus,11a The selectivity of these antiviral agents is due in part to the fact that they are appreciably I phosphorylated only in virus-infected cells, where a vi- OAC oh rus-specified thymidine kinase of low substrate specificity converts the nucleoside analogues to 5’-monophosphates. 6 X=H,R=ac 7 8 X=h The monophosphates are next converted to diphasphates 9 X=F. R:Ac 10 X=F and then to the corresponding nucleoside triphosphates by cellular enzymes. The triphosphates prevent virus 12 X=Br. r=h replication by inhibition of the viral DNA polymerase. Additional selectivity is realized at this stage because the our somewhat differing approaches to the recently reported host polymerase is less sensitive than the viral polymerase analogues. Additionally, we present new in vitro and in to the nucleoside triphosphate analogue,6b The potent and broad activity of DHPG against herpes viruses prompted us to synthesize other members of this (1) ContributionNo. 184 from the lnstitute of bio-organic Chem- class of acyclic deoxynuclesides In this report we describe istry, the synthesis of a number of pyrimidine and purine ana- (2) Presented in part et the 185th National Meeting of the Am- erican cehmical Society, Seattle, WA; carb 43; march 24. logues of DHPG, some of which were recently disclosed 1983. by Ogilvie and co-workers in a series of publications.12 (3) The structural formulas of DHPG (1) and the related acyclic This paper details the synthesis of new compounds and nucleoside analogues have been depicted in a ribose-like conformation only to draw attention to the similarity in . structure between these compounds and I
J. Med. chem 1986,28,347-368 347 A Reductase Inhibitors. 1. Structural Modification of 5-Substituted 3,5-Dihydroxypentanoic Acids and Their Lactone Derivatives
G. E. Stokker W. F. Hoffman A. W. Alberts E. J. Cragoe, Jr.? A. A. Deana,J. L. gilifillan J. W. Huff,' F. C. Novello, J. D. Prugh R. L. Smith,' and A. K. Willard Med. Sharp & Dohme, Research laboratories West Point, Pennsylvania 19486 and Rahway. New Jersey 07065. Received June 21, 1984
Formation of the atheromatous plaque or atheroma is acid (I), the dihydroxy acid form of mevinolin is the most accompanied by the locatized deposition of plasma lipids, potent HMG-CoA reductase inhibitor K 0.6 nM) re- primarily cholesteryl esters, in the intima of the arterial ported to date? Of even greater interest am the findings wall Growth of the atheroma eventually leads to con- that compactin and mevinolin9,13 are highly effective striction of the coronary arterial lumen and ultimately hypocbolesterolemic agents in several animal species and results in Arteriosclerosis and coronary heart disease (CH- man. Subsequent to the first reports disclosing the D), the major cause of death and disability in Western structure14 and biological activity of compactin a series countries these observation coupled with the compelling of studies directed tawad the developmentof structurally epidemiological eviidence implicating hypercholesterolemia simplified HMG-CoA reductase inhibitors were initiated as a primary risk factor for CHD,3,4 have stimulated re in these Laboratories. Described in this paper are the search on the development of therapeutic agents for pre- results of our initial study which served to delineate key venting and treating Arteriosclerosisbased on the attenu- ation of plasma cholesterol levels.5 The results of the recently completed Lipid Research Clinics Coronary (1) R Annu Rev. Med. 1979,30. I. Prevention Trial (LRC-CPPT)6 provide strong ross Primary (2) fuster Scand. Y. Haemotol. (Suppl. support for the basis of approach. the V. 1981,27 38),1. this LRC-CPPT (3) U.S. Department of Health and Human Services, National clearly demonstrated that reduction of low-density lipo- institute of Health, National Heart. lung and Blood insti- protein cholesterol (LDL-C) through dietary modification tute Arteriosclerosis VoL 1 and 2, nih Publication No. and treatment with the bile acid sequesterant cholestyr- 81-2034 and 81-2035, u.s. G.P.O., washington DC, 1981. amine, either alone or in combination, diminished the (4 Hamburg, D. A; Elliott, g. R Arteriosclerosis 1982,2,357. incidence of CHD morbidity and mortality in hypercho- (6) prugh J. D.; Rooney. C. S.; Smith, R L Annu. Rep. Med. lesteerolemic men at high risk for CHD. Nevertheless,the Chem. 1983,18,161. reduction of dietary cholesterol and saturated fat intake (6) (a) LRC-CPPT. J. Am. med assoc. 1984,251,351. (b) LRC- and the use of bile acid sequesterants often fail to lower CPPT. Ibid. 1984,251,386. elevated plasma LDL-C levels to the desired extent par- (7) Havel, R J.; kane J. P. Annu. Rev. med 1982,33,417. ticularly in patients with familial hypercholesterolemia (8) Endo, b; Kuroda M.; Taujita Y. J. Antibiot, 1976 29, 1346. (FH).7 (9) Alberts A W.;chen J; kuron G.; Hunt, V.; Huff, J.; hoff An attractive and potentially more efficaciousway to man C.; Rothrock, J.; lopez m joshua H.; Harris, E.; lower plasma cholesterol levelswould be to control de novo Patchett, A;Monaghan, R; currie S Stapley, E.; AIbers- cholestetogenesis by selectively inhibiting an early bio- Schonberg, G.; hensens 0.; Hirshfield, J.; Hoogsteen, X.; Liesch, J.; Springer, J. proc. Natl. Acad. sci. U.S.A. 1980,77, synthetic step. The highly functionalized fungal metab- 3967. olites compactin (ML-236B, CS-500)8 and mevinolin (10) rodwekk V. W.; Nordstrom J. L;Mitschelen, Y. J. Ado. Lipid (MK-803)9are potent inhibitors of cholesterol biosynthesis Rea. 1976, 14, 1. at the level of the major rate-limiting enzyme 3-hydroxy- (11) (a) Tsujita, Y; kurods m Tanzawa K;kitnao N.;Endo, A. 3-methylglutaryl-coenyme A reductase [HMG-CoA re- Arteriosclerosis (Shannon irel 1978 32,307, (b) kuroda M.; ductase; mevalonate:NADP oxidoreductase (CoA acy- Tsujita, Y.; tanzawa k Endo, a lipids 1979,14,585. lating), EC 1.1,1.34],10 which catalyzes the conversion of (12) (a) Yamamoto. A; Sudo. H.; Endo.A. Arteriosclerosis shan- HMG-CoA to mevalonic acid (eq 1). Indeed, mevinolinic non irel 1980,35,269. (b) Mabuchi, H.; haba T.; Tatami. R.; Miyamoto, S.; sakai Y.; wakasugi T,;Watanabe, I; ch3 koizumi J.; Takeda, R N. Engl. J. Med. 1981,306,478, (13) (a) tobert J. A; hitsenberger G; Kukovetz W. R.; Holmes, l B.; Jones, K. H. Arteriosclerosis (Shannon irel 1982,41, 61. (b) tobert d. abell G. D.; Birtwell, J.; james Iku- I mevalonic acid kovetz. W. R, Pryor, J. 5.; Buntinx A; Holmes, I. B,;chao SCoA Y.S.; bologness J. A. J. Clin. invest 1982,69,913. HMG-CoA (14) The X-ray structure of compactin was first reported by Brown. A, G.; Smale, T. C.; king T. J.: hansenkamp R; Thompson R H. J. Cham. soc.Perkin trans 1 1976,1165. merck Sharp & Dohme, west Point, PA Nota that the relative configuration in Figure 1 of the cited Merck Sharp & Dohme, Rahway, NJ. reference does not agree with the srystal coordinates we present address Stuart Pharmaceticals Division of IC1 present hem the correct relative and absolute stereochemical americas Wilmington, DE 19897. configuration of compactin 348 journal of Medicinal chemistry 1985 Vol. 28, No. 3 stokker et al SARs for compactin-like mimics an, afforded a series of alightly modified of Noltes et al.17 provided moderately effective HMG-CoA reductase inhibitors typ- 4-hydroxy ketone 11 in high yield. Acylation of 11 with typified by the ring-opened form of lactone 6a 2-bromoacetyl bromide in the presence of pyridine fur- HO nished bromo acetate 12 which was ring closed to 4- hydroxy-4-methyl lactone 13 via an intramolecular Re- formatsky reaction." Substitution of triethylamine for pyridine in the acylation step resulted in elimination of the 2-bromoacetoxy moiety and isolation of the resultant dienone. an alternate route to lactone 13 starting from 4-hydroxy ketone 11 was investigated This mute involved sequential acylation, intermolecular Reformatsky reaction with ethyl 2-bromoacetate, basic hydrolysis, acidification, and lac- tonization This route was abandoned in favor of the more efficient two-step route (vide supra). Ethylene lactone 13a was reduced in the same manner as 6a to provide the corresponding ethyl-bridged compound 16. Refluxing a solution of 13a and toluene in the presence of pTSA (trace) resulted in the smooth conversion of 13a to 17. 5-Methoxy-3-hydroxyheptenoic acid 20 was prepared by treating the dimethyl acetal of 2 with diketene (1 equiv) in the presence of TiC4 by using the general procedure CI of izawa and Mukaiyama19followed by borohydride re- 6a duction, basic hydrolysis and acidification of the resultant 3-keto ester 19 as shown in Scheme III. A similar con- Chemistry. The compounds prepared for this study densation of aldehyde 2 provided a mixture of 3-keto ester are listed in Tables I-III Their syntheses from the 3 and dihydro lactone 4 and, thus, was a less expeditious corresponding aldehydes, exemplified by 2, are shown in route to target lactone 6 than was the dianion procedure Schemes I-III Condensation of aldehyde 2 with the (Scheme I). dianion of ethyl acebacetate16 followed by borohydride The previously undescribed requisite aldehydes were reduction, basic hydrolysis, acidification and azeotropic prepared as shown in Schemes lV-VIII. The synthesia removal of water provided a mixture of the trans (64 of the unsaturated aldehyde 55a needed for elaborating and cis (6blactones which subsequently was separated lactone 55 is shown in Scheme IV with phenanthrene-4- by chromatography (Scheme i The use of MeOH in the carboxaldehyde as starting material This procedure was borohydride reduction step was found to be advantageous: also used to prepare the known unsaturated aldehyde replacement of MeOH by EtOH produced some of the precursors for lactones 56-58. corresponding ethyl ester which was more resistant to The 3-(decahydronaphthyl)propanals 25 and 27 (pre- hydrolysis. The resolution of 6a was accomplished via cursors to 51 and 52) were elaborated from 21 as shown formation and chromatographic separation of the diaste- in Scheme V. After high-pressure hydrogenation of 22, the reomeric (R)-a-methylbenzylamines followed by basic acid 23 was converted to aldehyde 25 by the Burgstahler hydrolysis and relactonization to yield 6a and 6a modification of the Rosenmund reduction20 The ethyl Hydrolysis and acidification of 5-hydrory-3-keto ester ester of 23 (24, isolated in about an equal amount during 3 without prior reduction resulted in spontaneous lacton- the workup of 23) was isomerized with AICl3 (2 equiv) at lactonization to enol lactone 4. Numerous attempts to reduce room temperature and subsequently hydrolyzed to acid 4 to hydroxy lactone 6, either catalytically or via metal 26, which was converted to aldehyde 27 in the same hydrides, were unsuccessful. Treatment of lactone 6a manner used for 23 25. with NH3 provided the erythro amide 10. Catalytic re- The Dibal reduction of nitriles 29,32, and 34 (Scheme reduction of lactone 6aprovided Compound 7 containing VI) provided the aldehydes requisite for preparing lactones a saturated bridging unit. The lactol ethers 8 and 9 were 47, 48, and 53, respectively. Aldehyde 37 (precursor to prepared by diisobutylaluminum hydride (Dibal) reduction 44 was prepared by alkylation of phenol 36 with the of 6a followed by treatment with MeOH in the pres- diethyl acetal of 2-bromoacetaldehyde followed by hy- ence of pyridinium p-toluenesulfonate (PPTS). drolysis (Scheme VII). The. syntheses of methyl lactones 13, 16, and 17 are finally conversion of cinnamaldehyde 2 to propargyl- illustrated in Scheme II A tin-mediated aldol condensa- aldehyde 41 (precursor to 42) was effected via the four step tion of aldehyde 2 with 2-acetoxypropene following a sequence 2 - 38 - 39 - 40 - 41 by using the general method of Allen and Edens22 as shown in Scheme VIII. (15) During the course of this study a series of mevalonolactone derivatives of the general structure were reported to inhibit. (17) noltes J. G.; varbeek F.; creemers H. m J. C. Organometal. Chem. Synth. 1970-1971. 1, 57. In the present case, (tri- butylstannyl)acetone was prepared in situ (18) For a general method, see maruoka K.; Hashimoto, S.: Ki- tagawa, Y.; Yamamoto, H.; Nozaki, H. J. Am. Chem. soc. 1977, 99,7705. (19) izawa T.; Mukaiyama, T. chem Lett. 1976,161. (20) Burgstahler, A. W.; Weigel, L. 0.;Shaefer, C. C. Synthesis 1976.767. HMG-CoA reductase by Sato et al.: Sato, A; Ogiso A.; no- (21) During the course of this investigation, the 4r,6r dechloro guchi H.;Mitsui, S.; kaneko L, Shimada Y. Chem. Pharm. analogue was prepared from tri-O-acetyl-D-glucal an a possible Bull. 1980,28, 1509. HMC-CoA reductase inhibitor by Yang. et d.: Yang. Y. L.; (16) Huckin. S. N.; Weiler, L. Tetrahedron Lett. 1971, 4835. Falck, J. R Tetrahedron Lett. 1982,23,4305. HMC-CoA Reductase Inhibitors journal of me chemistry 1985 Vol 28 No. 3 349 table I. Effects of lactone Modification urd Stereochemistry cl .
no. R EtOAc 62 168-169 2 0 10 12 50 15 acetonehexane 33 148-160 1 13 5 41 10 64 20 & acetone/hexane 17 116-117 2 4 5 6 10 27 20 17 50 18 n-BuC1 25 114-116 1 26 2 45 5 66 10 72 20 86 n-BuC1 44 114-116 1 0 2 0 4 0 8 0 chromat acetone/CH2Cl2 33 wax 2 0 4 2 6 4 chromat. acetone/CH2CI2 64 88-93 2 0 4 0 8 6 10 0 acetone/hexane 63 117-118 1 0 2 0 4 0 8 0 n-BuC1 35 138-138 1 11 2 21 4 36 10 61 n-BuCl/hexane 5.4 135-137 2 3 4 7 10 5 25 15 chromat CHCl3/MeOH 90 108-110 2 0 4 3 10 3 25 5 chromat Ch2CI2/HOAc 42 gum 1 0 2 0 5 0 10 0 analytical results are within 0.4% of the theoretical values unless otherwise noted- see Experimental section for protocol tested in the form indicated since carboxylate anion could not be formed under this testing protocol pK 5.22 (30% EtOH). 'When tested in the lactone form only 25% inhibition was observed at 50 µg/ml 'Yield from 12. about a 43 ratio of erythro and threo- overall yield from 18. anal Calcd: C. 52.68. Found C, 52.02.
The synthesiaof 43 (the Z isomer of 6a was accom- hydroxycarboxylate forms of each compound were tested accomplished by the catalytic hydrogenation-- (Lindlar) of 42. for intrinsic inhibitory activity. In each instance, the so- Biological results and Discussion dim dihydroxycarboxylate formproved more active than the lactone form see Table I, footnote e). Accordingly, The compounds in I-IV were evaluated for subsequent tests were done exclusively on the sodium their ability to inhibit solubilized, partially purified rat dihydroxycarboxylate forms unless otherwise liver HMG-CoA reductase. During the initial phase of this The contributions of lactone moiety Stereochemistry and study both the lactone and the ring-opened sodium di- functionality to intrinsic inhibitory in compound 6 are illustrated in table I. Separationof the lactone 22 Allen, C. F. HEdens C. OJr. organic syntheses wiley mixture 6 into the racemic cis 6band trans 6a New york 1955 collect VoL III isomers showed that activity resided principally in the 7 ch2ch2 n-BuCl 60 96-98 1 29 2 38 6 57 10 80 chromat. CH2tC12/acetone 50 87-88 1 14 2 26 4 37 8 63 42 c=c 0.625 0 3.125 8 43 0.625 0 3.126 0 44 OCH2 1 21 2 24 4 18 10 21 45 n-BuC1 13d 133-136 6 4 10 26 20 27 50 23
46 16 9 7 18 47 ch2ch2 19 29 45 61 chromat. CH2Cl2/acetone 16d Oil 11 7 25 36 Analytical results are within 0.4% of the theoretical values unless otherwise noted See experimental Section for protocol equatorial 4-Me in lactone by reduction of 13a overall yield from aldehyde. HPLC purification on Dupont silica 10/30 with i- PrOH-hexane (1:19, v/v) at 2 mL/min. times of elution are 13.2 min for 42 and 21.8 min for 43 racemic trans lactone 6a Resolution of 6aaf- methyl ether (20). These results demonstrate the im- forded enantiomers 6a and 6a(-);their evaluation portant contributions of the carboxylate and 5-hydroxyl showed that the activity displayed by the raacement re- groups to activity. finally it should be noted that lactol sulted solely from the dextrorotatory isomer The addition ethers 8 and 9 displayed greatly diminished activities. of a methyl group tothe 4-position of 6a to give trans The effects of altering the moiety bridging the aromatic lactone 13a, a compound which more closleyresembles the and the lactonefragments in 6aon intrinsic inhibitory HMG moiety of the substrate HMG-CoA, did not alter activity are shown in Table II. saturation of the ethenyl activity appreciably. However, cis lactone 13b, which bridge in 6aand ita 4-methyl derivative 13agave possesses the opposite relative stereochemistry at C-4, was ethyl-bridgedcompounds 7 and 16, respectively with little much leas active as anticipated from the results obtained change in activity. However, other modifications of the for 6a and 6b interestingly oxidation of the 4 bridge such as replacement with the ethynyl 42 cis- hydroxyl group of 6a to provide enol 4 greatly reduced ethenyl(43 and oxymethylene(44) group resulted in loss activity. This result is a likely consequence of the fact that of activity as did complete removal (45) of the bridging 4 readily forms the sodium salt of the enolate and moiety. In a companion series of naphthalene andogues therefore, fails to undergo ring opening to afford the re- (Table II compound 47 containing the saturated two- quired carboxylate anion under alkaline condition+ Re- carbon bridge proved superior. Increasing the length of placement of the enolic hydroxyl group in 4 with a methyl the bridge to three carbons (48) reduced activity and group to provide 17 further reduced activity" Replacement elimination of the bridge to provide 46 further reduced of the carboxyl group in the ring-opened form of 6a activity. with a carboxamido group (10) ablated activity as did The results of various carbocyclic moieties substituted conversion of the 5-hydroxl group to the corresponding at the 6-position of the lactone ring are shown in Table Table III Effects of 6-Substitution
Et2O/pet. ether 8-10 66-68 6 1 10 0 20 6 50 36 Et2O/ hexane 8-10 69-70.5 5 23 10 6 20 17 50 34 ch2ch2 Et2O/hexane 8-10 68-71 5 15 10 22 20 44 50 70 Et2O/ hexane 8-10 102-110 1 1 5 13 12.5 28 Et2O/hexane 8-10 122-129 1 18 5 16 12.5 28 Et2O/ 23 90-98 5 3 10 16 20 14 50 31 n-BuC1 25 140-142 0.36 39 0.75 58 1.0 64
n-BuC1 3 151-152 0.1 25 0.2 48 0.4 61 0.8 81
n-BuCI 12 gum 0.1 20 0.2 34 0.4 42
n-BuC1 10 143-144 0.1 22 0.2 36 0.4 14 0.8 20
overall yield from aldehyde. Analytical results are within 0.4% of the theoretical values unless otherwise noted. see Experimental Section for protocol mixture of trans/cis; 1.4/1. percent inhibition calculated an If contribution of cia isomer was zero anal Calcd: C, 68.99. Found: C, 68.54. mixture of trans/cis; 28/1. Anal Calcd: C, 72.82. Found C. 72.27. 'Mixture of trans/cis; 3.8/1. 'Mixture of trans/cis; 2/1. preparation of aldehyde, mp 112-116 "C;Hennion, G F.; Fleck, B. R J. Am. Chem. Soc. 1955, 77,3253. 'Preparation of aldehyde, mp 96-98 °C bergmann E. D.; Weiler-Feilchenfeld, H.; Mandel. N. Vietnamica Chim. Acta 1966, 129 Chem. Abstr. 1972, 72. 3276s. Preparation of aldehyde. mp 42-44 °C Kohler, E. P.; Larsen R. G. Y. Am. Chem. Soc. 1935,57, 1452. III The decahydronaphthalenes 51 and 52 and the ada- binding to HMG-CoA reductase is sensitive (a) to the mantyl compound 53 all possessed similar activity, while stereochemistry of the lactone moiety, (b) to the ability the cyclohexanes 49 and 50 were less active. Aromatization of the lactone moiety to be opened to a dihydroxy acid, of the cyclohexane ring to give the phenylderivative 54 c to the length of the moiety bridging the lactone and had little effect on activity. However, substitution of the the lipophilic groups, and (d) to the size and shape of the bridging moiety with larger aromatic groups such as those lipophilic group. Further modifications of the lipophilic in compounds 55-57 increased activity about 10-folds i.e., group leading to more potent inhibitors will be described to about 1% of the inhibitory activity of compactin. in subsequent papers from these Laboratories. Scissionof the 4a-4b bond of compound 56 provided a less Experimental Section compact molecule 58 with diminished activity. and relative values the most active Melting points were determined on a Thomas-Hoover capillary The IC50 potency of melting point apparatus and are uncorrected Proton NMR compounds evaluated in this study are compared in Table spectra were recorded in CDCl3 unless noted otherwise, on either IV. Although the compounds described above are only a Varian T-60, EM-390,or NT-360spectrometer Chemical shifts moderately active HMG-CoA reductase inhibitors, analysis are reported in parts per million relative to Me,Si as the internal of their intrinsic inhibitory activities suggests that inhibitor standard Elemental analysis for carbon hydrogen, and nitrogen stokker et al scheme I OH
CI 0 Cl 2 4
3
10 9 a CH2COCH2CO2CH3 H*. OH'. NaBH,, EtOH. C6H5CH3 A. H,, Rh/C g Dibal, h CH3OH. PPTS. NH,. (R)-(+
Table IV. In Vitro Inhibitory Potencies against HMG-CoA (0.125 g) in CH3oH (2 mL) was added dropwise to a stirred Reductase suspension of (7.5 g. 0.043 mol) in ac- etaldehyde (30mL) cooled in an ice bath. The resulting solution IC50.a,b rel was stirred 30 min with cooling diluted with acetic anhydride no. µM potency. (25 mL), and heated at 120 °C for 1 h This mixture was cooled compactin 0.01 100 diluted with H2O (60 mL) and 6 N HCl(25 mL), and heated at 6a 22 0.08 100 °Cfor 0.6 h. The light brown, oily product solidified upon 6a 10.8 0.16 cooling It was collected dried, and triturated with Et2O to provide 7 15.2 2 8.5 g, 98%mp 106-108 'C. An analytical sample wan prepared 13a 20 0.09 by recryetallization from hexane to provide 2 as a pale yellow solid: 16 19.8 0.08 107-108 6,70 dd, J = 15,6 7.20-7.73 (3 47 129 mp "C; NMR (H, Hz), H, 7.87 d. J 9.8 d, J = And 51 107 m), (H, 15 Hz), (H, 6 Hz). 52 90 (C9H6CI2O) c, H. 55 1.9 1.1 Methyl 56 0.89 1.5 heptenoate (3) wan prepared by a modification of the procedure 57 1.6 0.9 of Weiler,16 Methyl acetoacetate (23.2 g, 0.2 mol) was added dropwise to a stirred suspension of sodium hydride (50% oil 'Relative precision is 10% bIC50 is the micromolar concen- suspension) (10.5 g, 0,22 mol) in anhydrous (500 at 0 tration of the inhibitor required to give 50% inhibition under the THF mL) conditions of the assay system. °Cunder a N2atmosphere. The resulting solution was stirred 15 min at 0 °C and then treated with a 22 M solution (95.4 mL, 0.21 mol) of n-butyllithium in hexane over 10 min The yellow were determined with a Perkin-Elmer Model 240 elemental an- solution was stirred 15 min at 0 °C and then was treated with alyzer and are within 0.4% of theory unless noted otherwise a solution of 2 (44.2 g, 0.22 mol) in anhydrous THF (250 ml Optical rotations were determined with a Perkin-Elmer Model The resulting orange solution was stirred 15 min at 0 °C and then 141 polarimeter. All starting materials were commercially available quenched by dropwise addition of 12 N HCI (48 ml The reaction unless indicated otherwise. mixture was diluted with H2O (300 mL) and extracted with Et2O (2) was prepared by (3 x 300 d).The organic extracted were combined, washed with modification of the procedure of Baker." A solution of NaOH brine (2 x 200 ml dried over MgSO,, and filtered. The filtrate wan evaporated in vacuo leaving a oil The red oil was stirred in petroleum ether (200 mL) in order to remove the mineral oil (23) baker B. R.; Janson, E. E.; Vermeulen, N. M. J. J. Med. The mixture was cooled and the petroleum ether decanted to chem 1969,12,898. provide 628 g (90%)of 3 NMR 2.83 (2 H,d, J 6 Hz),3.47 scheme II CI on o
CI 11
Ac
12 14
\ CI 20 CI 0 MeOH, PTSA. b TiCI4 C MeOH. NaBH4 OH-. \ H Scheme IV CHO \ 15 00
a b gel + cis isomer LiCCH=CHOC2H5 Silica CI 13b 13a Scheme V CO2H CHO I
16 (n-Bu)3SnOCH3 CH2=C(OAc)CH b HO2CCO2R. BrCH2COBr C5H5N. Zn, CuBr, Et2AlCl Ac2O C5H5N. BrCH2CO2Et OH-. h H+. C4H4CH3 H,, Rh/C. PTSA, C4H3CH3 24 23 25 (2 H,a), 3.70 (3 H,s 4.76 H m), 6.13 (H, dd, J 15,6 Hz),6.90 (H,d, J 15 Hz),7.0-7.5 (3 H, m). 4 The ester 3 (20 g, 6.3 mmol was stirred in 0.1 N NaOH (200 for 4 h The resulting solution was acidified with 6 N HCl to provide a yellow solid which was recrystallized to analytical purity: yield 0.93 g NMR (MegSO-d6 H H (2 m). 5.00 5.13 m), 4.67 dd, J = 15, 6 2.60 W, (H.s (H, (H, 26 27 Hz),6.97 (H, d. J 15 Hz 7.27-7.87 (3 H. m). 11.5 (H,bra). Methyl a CH2(CO2H29C5H5N. H,, Ru/C, EtOH. SOCl2 heptenoate 5 sodium borohydride (1.3 g, 33.7 mmol was H,, Pd/C, 2,6-(Me)2C5H3N. e AICI,. OH-. W+. added with stirring to a cooled solution (6 "C) of 3 (10.7 g, 33.7 mmol) in EtOH 100 mL) at a rate sufficient to maintain the with CH2Cl2-CH3OH 49:1 v/v. 500 mL) provided a forerun which internal temperature at 15-20 "C. The resulting solution was was discarded Continued elution with the same eluant (3500 mL) stirred an additional 2 h with ice-bath cooling and then acidified provided 5 as a yellow oil; NMR 6 1.60-1.93 (2 H,m), 2.50 (2 H. with 6 N HCl. The resulting mixture was diluted with H2O (250 d, J 6 Hz),3.67 (3 H. s4.13-4.77 (2 H. m), 5.93-6.40 (H.m), mL) and extracted with Et2O (3 X 200 mL The Et2O extracts 6.93 (H. d, J 16 Hz),7.17-7.50 (3 H, m). Anal. (C14H14Cl2O4 werecombined, washed with brine driedover MgSO and filtered H; C: calcd 62.68; found, 52.25. The filtrate was evaporated in vacuo to provide a yellow oil (10.4 g, 97% A portion ofthe oil was purified by medium-pressure 6aand 6bAn EtOH so- chromatagraphy on a 25 X 1000 mm silical gel column. Elution lution 100 mL) containing 5 (8.4 g, 26.3 mmol) and 1 N NaOH 354 journal of Medicinal chemistry 5, vol. 28 No. 3 SchemeVI organic extracts washed with brine dried MgSO4 and thefiltrate was evaporated in vacuo to providea mixture cn cn of acid and lactone 7.8g, 97% A solution of this mixture in toluene 100mL) washeated at reflux in a dean-stark apparatus After2 hthe dean-stark apparatus was replaced with a soxhlet containing 3-A molecular sieves 100 g). the solution was refluxed for an additional 4 h and then the toluene wan removed in vacuo leaving a yellow oil 7.2 g, 96%) which was a mixture of 6a and 6b The oil was chroamtographedon 0 silica gel column 500 g Elution with CH2Cl2-acetone 4:1 v/v; 900 mL) provided a forerun which was discarded Continued elution with the same eluant (300 ml gave the trans isomer 6a(2.5 g Recrys- tallization of the solid provided an analytical sample as colorless needless NMR acetone-dg 6 2.06 (2 H, m). 2.69 2 H,m). 4.43 (H.m), 5.42 (H,m), 6.49 (H.dd, J 15, 6 Hz),7.08 (H.d, J = 16 hz 7.33-7.59 (2 H, m), 7.79 (H,d, J 8 Hz). An isomeric purity of 99.8% was determined for 6a by HPLC on a 31 32 Whatman Partinil-5 RAC column with 15% 2-propanol/hexane as the eluant. The time of elution was 4.96 min at 8 flow rata of 6 mL/min. Further elution of the column with the same eluant 600 mL) gave the cis isomer 6b as a solid 1.25 g). Recrystallization gave an analytical sampleas colorless needless NMR acetone-dg 34 35 1.50-2.93 (4 H,m), 4.36 (H.m), 5.02 (H,m). 6.37 (H,dd, J 16,6 7.02 d, J 7.16-7.50 (2 m), 7.67 d. NCCH2CO2H, NH4OAc, C5H5N. b Dibal. Hz), (H, IS Hz), H, (H, NaCN. J 8 Hz). An isomeric Purity of99.3% was determined for 6b by HPLC on a Whatman Partisil-5 RAC column with 15% 2- Scheme VII propanol/hexane as the eluant The time of elution was 5.79 min at a flow rate of 6 CI ml/min CI trans (7). A solutionof 6a (1.6 g 5.2 mmol in thf (100 mL) was stirred magnetically and hydro- genated at room temperature under atmosphericpressure in the presence of5% rhodiumon carbon 150mg until 1.25molar equiv 36 37 of hydrogenhad been consumed After removal of the catalyst BrCH3CH(OEt)2. 2N by filtration, the filtrate was evaporated in vacuo, leaving a solid. The solid wan recrystallized to provide 7 (0.9 g): NMR 6 1.67-217 Scheme VIII (4 H. m), 2.60-3.13 (4 H,m). 4.30-4.50 (H,m), 4.57-4.90 (H.m), 7.14-7.44 (3 H, m). CI Resolution of (Sa). A solution of 6a 2.87 g. 10 mmol in (R)-(+)-u-methyl- benzylamine (15 mL) was stirred for 18 h at ambient temperature and then poured into H2o 100mL). This aqueousmixture was acidified with 6 N HCI and extracted with et2o (3 X 100 d). CI The Et2O extracts were combined washed with brine, dried over MgSO4 and evaporation ofthe filtrate in vacuo provided thecrude diastereomeric amides as a tan viscous oil (4.1 g. 100%). CI This oil (3.1 g, 7.6 mmol was chroamtographedon a silica gel column 200 g Elution with acetone-CH2Cl2 1:4v/v. 1200 ml gave a forerun which was discarded Continued elution with the same eluant provided the mixture of diastereomericamides as a viscous oil (3.0 g, 97%). This mixture was separated by chro- matography on a Waters Prep LC 500 The separation was and eluting with acetone-CH2Cl2 1:4 v/v). use of the shave- recycle technique provided diastereomer A (1.36 g) and diastereomeric reomer B (1.2 g). Recrystallizationof diastereomer A from n-butyl chloride gave colorless clusters (1.0 g) which melted at 106-108 'C; NMR 1.47 (3 H. d, J 6 Hz),1.70 (2 H, m), 2.33 (2 H, d. J 6 Hz),4.30 (H,m), 4.58 (H, m). 5.13 (H,m), 6.20 (H,dd. J 15,6 Hz), 6.33 (H,m), 6.93 (H,d, J 15 Hz),7.33 (8 H, m). Anal C21H23 Cl2NO2 C, H, N. Recrystallization of diastereomer B from n-butyl chloride gave a pale yellow didwhich melted at 55-60 °C NMR 1.47 (3 H, d. J 6 Hz), 1.70 (2 H, m). 2.33 (2 H, d, J 6 Hz), 4.33 (H,m), 4.60 (H,m), 5.16(H, m),6.17 (H. dd, j 15,6 Hz),6.23 (H.m), 6.93 (H.d, j = 15 Hz). 7.33 (8 H. m). Anal (C21H23C12NO3) C, 43 H,N. a Br,. K2CO (EtO)3CH9NH4CI KOH. Dilute 5% Pd/CaCO3 6adiastereomer A 0.74 g, 1.8 mmol was dissolved in 95% ethanol (25 mL) (26.3 mL) was stirred at ambient temperature for 1 h The containing 1 N NaOH (3.6 ml 3.6 mmol) and the solution was reaction solution was acidified with 6 N HCl diluted with H2o refluxed for 54 h The solvent was removed in vacuo leaving a 200 mL). and extracted with et2o (3 x 100 d).The combined residue which was mixed with H2o and acidified with 6 N HCl HMG CoA Reductase Inhibit on journal of inal chemistry 1985 Vol. 28 No. 3 355 the resulting mixturewas extracted with et2o,the et2oextracts H, d, J 6 Hz), 4.73 (H,m), 6.10 (H, dd And (C12H12Cl2O2 werecombined washed with brine,dried over MgSO4 and filtered C, H. The filtrate wan evaporated, leaving the intermediate acid as a 2-Bromoacetate yellow oil (0.54 g A solution of the oil in toluene (150 mL) was (12). 2-Bromoacetyl bromide (1.1 mL. 13.2 mmol) was added refluxed through a SoxhIet containing mol& sieves (3 A) for dropwise to astirred solution of 11 (3.4 g, 13.1 mmol and 5 h. The solution was evaporated in vacuo, laeving the lactone (1.07 mL, 13.2 mmol) in et2o (100 ml at 0 *C. The ice bath 6aas a yellow solid Recrystallization gave colorless needles was removed and the reaction mixture was stirred at 20 °C for (0.13 g): [a]25D 5.9 (c 0.426, CHCl3 NMR 203 (2 H, m), 273 2 h and then diluted with H2o (100 ml The organic layer was (2 H. m), 4.46 (H.m). 6.41 (H, m). 6.19 (H, dd, J 15,6 Hz).7.01 separated and washed with 1 N HCI (100 mlH2o (2 X 100 ml (H,d, J 15 Hz). 7.14-7.50 (3 H, m). andsaturated brine, dried (MgSO4),filtered, and evaporated the trans residual oil was chromatographed on silica gel. Elution with 6a Diastereomer CH2Cl2-acetone 99:1 v/v; 19 L) provided 12 (2.8 g, 56%): NMR B (1.1 g. 27 mmol wan dissolved in 1 N NaOH (5.4 mL, 5.4 mmol 6 2.2 (3 H, m), 2.92 (2 H,t, J 7 Hz),3.85 (2 H, a). 6.9 (H. m). and the solution was refluxed for 18 h. The same workup and 6.15 (H. m), 6.95-7.5 (4 H. m). Replacement of pyridine with an lactonization used in the synthesis of 6a gave 6a(-) as a yellow equivalent of TeA provided only the dienone (6(2-4dichloro- solid Recrystallization provided colorless needles (0.34 g mp as a yellow powder (77% mp 80-82 114-115 °C [a]25D -6.6° (c 0.655. CHCI3); NMR 6 2.03 (2 H. m), 'C; NMR 2.28 (3 H, s 6.15 (H,d, J 15 Hz),6.7-7.6 (6 H. 273 (2 H. m), 4.46 (H, m), 5.41 (H,m), 6.19 (H, dd. J 15,6 Hz). m). 7.01 (H, d, J 15 Hz), 7.14-7.50 (3 H, m). The optical purities of 6a and 6a were determined by NMR with ca. 0.5 molar (13). A solution of 12 (2.8 equiv of eu(hfbc)3 in CDCl3 each enantiomer was found to be g, 7.4 mmol) in dry THF (50 mL) was added dropwise to a vigorously free of the other enantiomer within the limits of detection orouslystirred slurry of activated Zn dust (720 mg. 11.1 mmol). (threshold ca 2%). Therefore, the optical purity of 6a and CuBr (60 mg, 0.4 mmol). et2alcl (25% solution in toluene; 3.2 6a was estimated to be 98 2%. mL, 8 mmol), and dry THF (50 mL) under N2at 20 "C. Stirring (2R4R 6S and (2S 2,4-Di- wan continued for 5 h before quenching with pyridine (8 mL) chlorophenyl)et followed by H2o (500mL) addition and et2o extraction (3 x 150 hydropyran (8 and 9). A solution of Dibal(0.89 m in hexane ml The combined et2o extracts were washed with 1 N HCl (4.31 mL, 3.83 mmol was added dropwise over 10 min to a stirred (2 x 50 ml H2o (2 x 250 ml and saturated brine, dried toluene (20mL) and THF (10 mL) solution of 6a(0.5 g, 1.74 (MgSO4),filtered and evaporated, leaving a sticky, pale yellow mmol) which was cooled to -78 "C. after stirring another 30 min solid (1.8 g) which was a mixture of 13a and 13b. This crude at -78 °C the reaction was quenched by the addition of MeOH productwas digested oncewith et2o (40 mland then crystallized (613 mg, 19.15 mmol). After the reaction mixture had warmed from n-butyl chloride (25 mL) to provide the trans isomer 13a to ambient temperature it was diluted with H2o (1 mltreated as tiny colorless crystals (550 mg): mp 136-138 °C NMR 6 1.4 with Celite, and stirred for an additional 15 min. Anhydrous (3 H. a, CH3),1.73 (5-Ha,dd, J 11, 13 Hz), 2.06 (5-H ddd, J Na2SO4 wan added and stirring was continued another 15 min. 2, 11,13 Hz), 2.12 (W.s OH), 2.45 3-Ha, d. J 18 Hz). 2.73 This mixture wan filtered and the filtrate evaporated in vacuo (3-H dd, J 2,18 Hz). 5.2-5.45 (6-H., m), 6.13 (H,dd. J 15, to provide a mixture of lactols (0.5 g, 99%). A CH3OH (10 mL) 6 Hz). 6.92 (H,d, J 15 Hz),7.67-7.43 (3 H. m). solution of the lactol mixture (0.25 g, 0.86 mmol) was treated with The filtrates from digestion and crystallization vide supra were PPTS (25 mg) and then refluxed through a Soxhlet containing combined, evaporated. and chromatographed with a Waters Prep molecular sieves (3 A). The reaction was cooled and diluted with LC 500. The separation was accomplished by useing two prep et2o (200 ml After washing with saturated NaHCO3 solution. PAK-500 silica cartridges in series and eluting with CH2CI2- H2o and brine, the organic layer was dried over MgSO4 and acetone (15:1, v/v). By use of the shave-recycle technique, the filtered. The filtrate wan evaporated in vacuo to provide the cis (b, 220 mg) and the trans a 230 mg) isomers of 13 were mixture of 8 and 9 (255 mg, 78%). separted The cia isomer (13b) wan crystallized from n-butyl this mixture was separated by flash chromatography an a silica chloride-hexane (2:1,v/v) (120 mg): mp 135-137 "C; NMR 1.46 column Elution with CH2CI2-acetons 19:1 v/v) first provided (3 H, CH3),1.8-2.4 (3 H. m,5-H OH). 2.65 (2H. a. 3-H the minor isomer 9 followed by the major isomer 8. 4.8-5.1 (6-Hm), 6.26 (H. dd, J 15,6 Hz). 7.03 (H.d. J 15 homer 9 NMR 6 1.47-2.13 (4 H. m), 3.44 (3 H, s 3.53 (H. Hz). m), 4.13 (H,m), 4.77 (H, m), 6.20 (H,dd. J 15,6 Hz). 6.98 (H. The epimeric alcohols were readily distinguished by analytical d, J 15 Hz). 7.13-7.57 (3 H, m). TLC (fluorescent silica gel (40 A), 1 x 3 in., MK6F, Whatman) isomer 8: NMR 6 1.47-1.93 (4 H, m), 3.53 (3 H, a), 4.40 (H. with CH2Cl2-acetone 9:1 v/v) as the eluant; R,0.25 for 13b and m), 4.63 (H, m), 4.85 (H,dd, J 9.3 Hz), 6.23 (H, dd, J 15, 0.30 for 13a 6 Hz), 6.98 (H.d. J 15 Hz),7.10-7.67 (3 H. m). Alternate Routs to 13 4-Acetory-6-(2,4-dichIoro- erythro- (14). Acetyl chloride (1.2 ml16.5 mmol) tenamide (10). lactone 6a(6.0 g, 20.9 mmol was dissolved was added dropwise to a stirred solution of 11 (3.9 g, 15 mmol in dry MeOH (100 mL) and saturated with anhydrous NH3 at in pyridine (60 ml cooled at 0°C. The ice bath was removed 0°C The reaction was sealed and stirred at room temperature and the reaction mixture was stirred at 20 °C for 2 h and then for 13 h, stored in the freezer overnight, and then worked up by diluted with et2o (300 ml The et2o solution was washed with evaporation under reduced pressure Two recrystallitionsfrom 1 N HCI (3 X 300 mland saturated NaHCO3, dried (MgSO4), acetone-hexane gave 10 (4.0 g): mp 117-118 'C; NMR 6 1.72 (2 filtered and evaporated, The residual pale amber oil (4.1 g) was H, m), 2.40 (2 H, d. j 6 Hz),4.22 (1 H, m), 4.55 (1 H, m),5.00 chromatographed on silica gel Elution with CH2Cl2 (2 L) provided 14 as a pale yellow oil (3.95 g, 87%); NMR 2.03 (3 H, s 2.17 Ha), 6.8-7.75 (3 H, m). (3 H,s 2.83 (2 H,dd, J 7,2 hz Anal. (C14H14Cl2O3 C, H. (1 1). Ethyl tri-n-butyltin methoxide 2-Acetoxypropene(3.3 mL, 30 mmol and tri-n-butyltin methoxide methyl-6-heptenoate (15). A solution of 14 (1.3 g, 4.3 mmol) (5.7 g, 24 mmol were combined and stirred at 60-70 "C under and ethyl bromoecetate(0.47 mL, 4.2 mmol in dry THF (10 mL) N2 for 1 h and then placed under vacuum for an additional 30 was added dropwise to a vigorouslystirred slurry of activated Zn min. Propenal 2 (4 g, 20 mmol) was added and the reaction dust (490 mg 7.5 mmol CuBr (29 mg,0.2 mmol et2alcl(25% mixture was stirred at 70 'C under N2 for 4 h. The dear reaction solution in toluene; 1.72 mL, 4.3 mmol), and dry THF (5 ml mixture was then cooled treated with malonic acid (1 g, 10 mmol) under N2 at 20 "C. Stirring was continued for 5 h before in et2o (20 ml and refluxed for 30 min. After cooling to -20 quenching with pyridine (3.5 mL) followed by H2o (50 mL) °C.the reaction mixture was filtered and the precipitate was addition and et2o extraction (3 x 80 ml The combined et2o washed with et2o (4 X 10 ml The et2o solutions were com- extracts were washed with 1 N HCI (2 X 25 ml H2o (2 X 50 bined and evaporated and the residual oil was chromatographed mland saturated brine dried (MgSO4), filtered and evaporated, on silica gel Elution with CHCI3-MeOH 99:1v/v 2 L) provided leaving crude 15 as a pale yellow oil (1.2 g, 75%): NMR 1.28 11 an a thick yellow oil (4.2 g, 81% NMR 2.2 (3 H, a). 2.73 (2 (3 H. t J 7 Hz 1.33 (3 HI), 2.10 (3 H, s 2.45 (2 H, d, J . 354 journal of medicinal Chemistry, 1985 vol 28 No. 3 stokker etal 13 H 4.16 (2 H, q, J 7 Hz Compound 13 this compound was prepared similary to 6. With 15 (1.2 g, 3.2 mmol as starting material 13 (120 mg) was at asteady rate and was complete in 1 h. the reactionmixture obtained as a mixture of trans 54%) and cia (46%) isomers as was filtered and the filtrate was evaporated. the residue was determined by HPLC Whatman partisil-10 PAC eluting with distributed between 6% HCI (200 mL) and Et2o 200ml the i-PrOH-hexane 1:10v/v) at 21 mL/min). Times of elutionare et2o was washed with aqueous \ H2O and saturated 3.0 min for a and 4.0 min for b brine, dried (MgSO4 filtered and evaporated theresidue was trans distilled in vacuo to provide 25 as a clearcolorless oil (27g,69.5%): 16 this compound bp 96-99 °C (0.5 mm); NMR 6 1.0-2.0 (19 H, m 2.50 (2 H,dt, prepared analogously to 7, starting with 13a 100mg, 0.33 mmol). anal (C13H22O)H;C: calcd The residual oil was chromatographed on silica gel Elution with 80.35 found, 79.70. CH2Cl2-acetone 19:1 v/v; 350 mL) provided 16 (50 mg,50%): 3-( trans-syn Acid NMR 1.37 (3 H, s 1.6-2.1(4 M m), 2.5-3.1 (4 H. m). 4.55-4.90 (26). Ethyl propanoate (24) 24 g, 100 mmol) was mixed with (H.m), 7.15-7.25 (2 H. m). 7.33-7.43 (H,m). anhydrous AlCl3 (27 g, 200 mmol at 0 °Cand then stirred at 20 (E6 'C for 70 h. The viscous reaction mixture was cooled to 0 "C, methyl-2H-pyran-2-one (17). A solution of 13. 150 mg, 0.5 quenched with ice-cold 1 N HCl(500 mL), and extracted with in toluene (75 mL) containing (5 was mmol PTSA mg) refluxed et2o 4 X 100 ml The et2o extracts were combined and through a Soxhlet filled with 3-A molecular siaves for 6 h After evaporated to afford a residue which was stirred with 1 N NaOH evaporation of the toluene, the residue wan chromatographed on (100 mL, 100 mmol) and MeOH (400 mL) for 2 h at 20 °C The silica gel Elution with CHCl3-MeOH 99:1V/V, 60 mL) provided reaction mixture was evaporated and the residue was distributed 17 (130 mg,90%): mp 108-110 'C; nmr 2.0 (3 H,s 2.4-2.6 between H2O (200 mL) and et2o (200 mL). The aqueous layer (2 H. m). 5.0-5.25 (H, m), 6.85-6.0 (H, m), 6.25 (H,dd, j = 16, was cooled to 0 'C and acidified with 12 N HCl to provide 26 as 6 Hz), 6.95-7.6 (4 H, m). a sticky yellow solid tion from petroleum ether gave Dimethyl Acetal (18). A 9.1 g 43% mp C13H22O2 c, h solution of 2 (10 g. 50 mmol), PTSA 50 mg). and anhydrous 3trans syn- (27). MeOH (100 mL) was refluxed through a Soxhlet filled with 3-A this compound was prepared similarlyto 25 starting with 26 (10.1 molecular sieves for 1 h. The solution was cooled treated with g, 48 mmol). The intermediate acid chloride was obtained as a anhydrous K2CO3 filtered and evaporated The residual oil was clear colorlessliquid (10.2 g, 93%): bp 113-116 °C 0.8-1.0 mm); chromatographed on silica geL Elution with CH2CI2provided 18 NMR 0.6-1.9 19 H, m), 2.88 (2 H,t, J 9 Hz). And C13 as a clear, pale brown oil (10.4 g, 84%). H21ClO) C, H. Reduction provided 27 (74%): bp 107-112 °C Methyl (1.3-1.5mm); NMR 0.6-1.8 (19 H,m), 2.43 (2 H,dt j 8,1 noate (19). A solution of ml mmol ticl4 in CH2Cl2 (3 M, 10 30 Hz). 9.85 (H, t J 1 Hz).anal (C13H22O) H C: calcd 80.35 was added dropwise to a vigorously stirred solution of 18 (6.18 found, 79.94. g, 25 mL) °C under A solution mmol in CH2CI2 (40 at -78 n2 (E)-3( (28). A mixture of of diketene in mL, 50 mmol) wan added in a CH2Cl2 (6 M, 10 (156g, 1.0 mol), cyanoacetic acid 78g, 0.9 mol), steady stream. The dark reaction mixture was stirred at -78 °C and ammonium acetate (3 g) in toluene (200 ml and pyridine for 1 h and then anhydrous MeOH 20 ml was added. after (100 mL) was heated at reflux for 2 days water wan collected in stirring for min 'C, the yellow solution an additional 30 et 20 light a Dean-stark trap. after evaporation of the solvents the product was poured into cold, aqueous K2CO3 (10 g/L). The yellow salts was distillation vacuo 140-185°C, 0.7 mm). were separated byfiltration andwashed withCh2Cl2 Theorganic obtained by in Recrystallization from EtOH-H2O gave 28 (145.7 g, 88%): mp layers were combined, washed saturated brine, with H2o and dried 69-71 'C; NMR 6 5.94 (1 H,d, CH,J 17 Hz),7.25-8.1 (7 H. (MgSO4and evaporated, leaving 19 as an orange oil (6.6 g, 80%) m,C10h7 8.25 (1 H,d, CH,J 17 Hz). anal (C13H9N C, H which exhibited one major spot (R 0.45 on tlc(fluorescent silica gel x 3 in MK6F, after eluting N. (40 A), 1 whatman with (29). A mixture 99:1 v/v): NMR 2.90(2 d, J 4 of 28 CH2Cl2-MeOH H. Hz),3.43 g. mol) and 6% Pd/C (1.0 g) EtOH (200 mL) was (3 e), (2 a). 3.83 (3 4.13-4.57 h 6.10 (H, dd, (17.9 0.1 in H, 3.63 H, H,I), m). hydrogenated a Parr apparatus after filtration and removal J = 6.83-7.43 in 16, 7 Hz), (4 H, m). of the solvent,the product was obtained distillation vacuo Acid (23) by in 'C, mm). recrystallization Et2o-petroleum and Ethyl ester 24. acid" (79.6 g, 150-165 0.5 from ether afforded 29 (16.4 g, mp 48-51 °C 2.72 (2 400 mmol) was hydrogenated EtOH mL) over 5% 90%): nmr in (600 Ru on t, J (2 H, t, J 8 7.25-8.1 (7 H, carbon (8.0 g) at 100 °C and 3000 psi- The catalyst was removed H, 8 Hz),3.4 Hz), m AnaL by filtration and the was evaporated residue wan (C13H11N) C, H, N. filtrate The 30 a stirred distributed between Et2o 700 mland 5% NaOH (2 x 400 mL). To suspension The et2o layer was washed with H2O and saturated brine, dried of 29 21.7 g,0.12 mol) in Et2o 400 mL) at -78 °C under n2 was (MgSO4 filtered and evaporated Distillation of the residue added a solution of dibal(85 mL, 0.128 mol) in toluene (25.3% 0.8446 g/mL) over a period after additional 1 at provided ethyl ester 24 as a clear colorless liquid (48.4 g, 50.7% of 1 h an h -78 ice-acetone bath was removed the reaction bp 120-122 °C(1.0 mm); NMR d 1.0-1.9 22 H, m), 2.27 (2 H, "C, the dry and t, J 6 Hz).4.14 (2 H, J 8 Hz). anal (C15H25O2 C. H. mixture was stirred at room temperature for 3 h The mixture The basic layer vide supra was extracted with (200 ml was added slowly to 5% aqueous H2So4 and then extracted with Et2o The solution was dried and evaporated and the and then acidified with 12 N HCI aftercooling to 0 'C. The acid et2o et2o was removed by filtration and from petroleum other residual oil distilled to give the product as a colorless oil bp to provide 23 as colorlessneedless (23.2 g, 27.6%): mp 80-82 °C 113-117 "C (0.2 mm which solidified on standing recrystal 0.8-1.8 (19 H, 233 (2 J 8 11.4 br I). lization from ether yielded 30 (16.8 g, 72%): mp NMR m), H, t Hz), (H, 282 (2 (2 J 8 721-8.02 anal (C13H22O2 C, H. 29-31 'C; NMR H,m), 3.4 W,t, Hz), 25 (7 H, m), 9.88 (1 H, t, J 1 hz anal (C13H12O) C, H. Propanoic acid 23 10g, 47.6 mmol) was dissolved in SOCl2 20 4 (32). A mixture of 3126 mL) and the solution was heated at reflux for 3.5 h The solution (58.8 g. 0.24 mol) and sodium cyanide (25 g, 0.6 mol) in EtOH wan evaporated and the residual oil was distilled in vacuo to (300 mland H2O (100 ml was heated at reflux with stirring provide the acid chloride as a clear colorless oil (7 g, 64%): bp for 5 h The reaction mixture was cooled concentrated in vacuo, 113-120 °C (0.8-1.0 mm); NMR 6 1.0-2.0 (19 H,m), 2.87 (2 H, and extracted with et2o "be etso solution was dried, filtered, t, J 8-Hz And. (C13H21C10C. h and concentrated to give the muds product. which was purified The acid chloride (4.56 g. 20 mmol) was added dropwise to a by distillation yield 38.8 g (84%); bp 151-155 °C (0.3 mm NMR well-stirred suspension of hydrogen-equilibrated 10% Pd/C 200
(25) UCLAF fr Patent 992 104 Chem. abstr. 1951.50, P15591f. (24) west B. L J. Am. Chem. Soc. 1920.42,1656. (26) rona P.; Feldman U. J. Chem. Soc. 1958 1737. HMG-CoA Reductase Inhibitors journal of med chemistry 1985 Vol. 28 No,3 357 1.7-2.3 4 H, m), 3.08 (2 Ht, J 7 hz 7.1-8.0 (7 H, m). Anal. andpurified through the second ammonium sulfate precipitation C, H, N. step u described by Kleinsek at al.34 The enzyme preparation 4-(1-Naphthalenyl)butanal (33). this product was prepared wastested forhmg-coa reductasepotency and diluted with 100 from 32 in a manner similar to that used for 30 and purified by mM phosphate buffer (pH 7.2) so that 100 µl of the enzyme distillation: bp 123-126 ºC(0.1 mm): NMR 213 (2 H. m), 2.50 solution when added to the assay control, gave a value of (2 H, m), 3.12 (2 H,t, J 8 hz 13-8.3 (7 H, m), 9.82 (1 H,t, 50000-60000 dpm. The enzyme preparation was stored at 80 J 1 Hz). ºC 35 prepared analogously to HMG-CoA Reductase Inhibition assay The assay is es- 33 starting with 3427 (4.73 g, 25 mmol Distillation under vacuum sentially the procedure of Shefer et al.36 The complete assay provided 35 as a Colorless oil (27 g, 56% bp 107-115 ºC 1.0-1.6 medium contained the following in a total volume of 0,8 mL mm) lit.28 bp 101-103 ºC (1.5 mm)];NMR 1.3-2.1 (17 H. m), phosphate Mer,pH 7.2,100 mM: MgCI2, 3 mM; NADP,3 mM; 2.4 (2 H. dt. J 9. 0.8 hz 9.85 (H,d, j 0.8 Hz). glucose 6-phosphate, 10 mM; glucose-6-phosphatedehydrogenase, (57) was prepared by 3 enzyme units reduced glutathione, 50 mM; HMG-CoA (glu- the method of Julia and tchernoff taryl-3-14C new England nuclear 0.2 mM (0.1 µci and partially (a) diethyl acetal purified enzyme stock solution 100 µl yield (45%);bp 140-142 ºC (22 mm) [lit.29 bp 137-140 ºC (2 test compounds or compactin after first being converted to mm)];NMR 1.23 (6 H, t. J 6 hz 3.43-3.93 (4 H, m), 4.0 (2 the dumsalt of their dihydroxy acid form in situ by addition H, d, J 4 Hz),4.80 (H, t, J 4 Hz),637-7.3 (3 H, m). of 1 N NaOH (1 equiv)) were dddto the assay system in 10-µl (b) 37 yield (80%);bp 125-128 ºC (2.2 mm) lit.29 bp 120 'C volumes at multiconcentration Ievels after a 40-min incubation (1.5 mm NMR 6 4.60 (2 H. a 6.67-7.4 (3 H. m), 9.8 (H,a). at 37 ºC with shakingand xposure toair, the reactionwas stopped was synthesized from by the addition of 0.4 mL of 8 N HCl after an additional 30-min 2 by the procedure used by Allen and Edens22 to convert 3- incubation period at 37 ºC to ensure the complete lactonization phenyl-2-propenal to of mevalonic add to mevalonolactone, 0.2 mL of the mixture was (a) (38): added to an 0.5 X 5.0 cm column containing 100-200-mesh bio- yield (67%). An analytical sample was crystallized from cyclo- Rex 5, chloride form bio-rad wetted with distilled water, as hexane-toluene: mp 125-126 'C; NMR 7.35-7.65 (2 H, m),8.2 described by Alberts et d.' The unreacted [14C]HMG-CoA was (H,d, J 6 Hz),8.25 (H, s), 9.55 (H,I). And absorbed on the resin and the 14C]mevalonolactone was eluted C, H. with distilled water (2 X 1 mL) directly into 7-mL scintillation (b) diethyl vials Five milliliters of Aquasol-2 (New England Nuclear) was acetal (39): yield (70%);bp 130-140 ºC (0.1-0.05 mm)as a pale added to each vial and radioactivity was measured in a Packard yellow oil; NMR 6 1.25 (6 H, t. J 6 hz 3.65 (4 H. dq, J 6, Tri Carb prias scintillation counter IC50 values were determined 3 Hz),5.0 (H, a). 7.2-7.75 (4 H. m). by plotting percentage inhibition against test compound con- (c) diethyl acetal (40): centration and fitting a striaght line to the resulting data by using yield (88%);bp 122-124 ºC (0.04 mm); NMR 6 1.23 (6 H, t, J the least-squares method. For estimation of relative inhibitory 6 N2), 3.5-3.95 (6 H, m), 5.5 (H,s 7,1-7,5 (3 H,m). Anal. potencies, compactin was assigned a value of 100 and the IC50 C H. value of the test compound was compared with that of compactin (d) 41: yield (67%). An analytical sample was crystallized from determined simultaneously. mp 'C; 7.2-7.7 (3 H, m), 9.5 (H, e). 57-58 NMR AnaL Acknowledgment. We are indebted to Drs. R. C, H. F. was prepared by Hirschmann and P. S. Anderson for their encouragement the general procedure of Wollenbeg.30 n-Butyllithium (2.72 throughout the course of this investigation, to Dr. W. C. mmol) was added dropwise to a solution of randall and staff for analytical support, to Drs. B. H. (2.3 g, 6 mmol) in dry THF (20 mL) arison and D. W. Cochran for NMR spectra, and to M. after stirring at -78 ºC for 1 h, a solution of 4- Z. Banker for manuscript preparation. °C (1.1 g, 5.3 mmol) in dry THF (10 registry NO. 2,70017-27-5: 4.93863- wan added rapidly the mixture was stirred at mL) and reaction 42-4 5,93863-436; 6a86097-37-2 6a78444-38-9; 6a -78 ºC for an additional 2 min before removing the cooling bath. 78444-39-0; 86097-383; 7,86097-55-4; after stirring at 22 ºC for 30 min,the reaction mixture was 6b 893863- treated 44-6 8(Iactol), 93863-81-1; -9,93922-58-8; 9(Lactol). with saturated NaHCO3 20 mL) and the reaction mixture wan 93922-80-2; 10 93863-46-8; 12 between (100 mL) (100 mL). et2o 938634&7; 11 distributed Et2o and H2o The 93363-47-9; 13a 9-3863-48-0 13b 93863-49-1: layer with h2o (3 x 100 mL), dried mgso4 filtered, was washed 93863-50-4 1583883-516; 17,93863-53-7; and then evaporated. Chromatography of the biphasic residue 18, 93863-54-8;19,93863-559; 93863-56-0; on silica gel with elution by CH2cI2 provided 55a (900 mg, 73%): 93863-67-1; 21. 66-77-3; 22, 13026-12-5; 23 mp 112-120 'C. Crystallization from hexane yielded analytically 93863-582; 23 acid chloride), 93863-59-3; pure 55a mp 125-129 'C; NMR 6.8 (H, dd, J 18, 8 Hz). 93863-62-8; 26 (acid chloride), (10 m), 9.9 d, J 8 C, H. 259386361-7; 26 7.5-8.45 H, (H. Hz). AnaL 93863-63-9 27,93863-64-0 28,93863-65-1;29,70067-70-8; Isolation of HMG-CoA Male Holtzman 30, Reductase. 53531-16-1; 31,27650-86-8; 32,93863-66-2;33,93863-67-3; 34, Sprague-Dawley rata (225-250 g) were kept on reversed lighting 52582-89.5; 36,18228-55-2;36, 120-83-2 37,17944-27-3;37 (diethyl chow 3% Cholestyraminfor 7 days and fed purina rat containing acetal), 78830-79-2 39,93863-69-5; 40,93863-70-8 preceding their asphyxiation Livers were 38,93863-684; sacrifice by CO2 re- 41, 93863-71-9; 93863-72-0; 43 93863-73-1; DL-44, moved h the cycle and used immediately to prepare 42 6 into dark 86097-50-9; 45 86097-49-8: 46 93863-74-2; 47, microsomes HMG-CoAreductase was solubilized from the freshly 93863-75-3; 48 93863.764; 49 50 prepared microsome by the method of Heller and Shrewsbury33 93863-77-5: 93922-69-9; 51 93863-786; 52 93863-79-7; (53, 93863-80-0; 54 86118-13-0 55 93863-81-1 55a (27) McKervey, M. A. J, chem soc Perkin trans I 1972, 2707 93863-82-2; 56 93863-834; 57 93863-84-4; 58 (28) stepanov F. N.: Dovgan, N, L Zh. Org. Khim. 1968,4,277; 93863455; BrCH2CH(OEt)22032-35-1; CH3CHO, 75-07-0; Chem. abstr 1968- 68,104612w CH2COCH2CO2CH3 105.45.3; 10822-5; (29) Julia, M;tchernoff G. bullsoc chim fr. 1953,479;Chem. CH2(CO2H)2141-82-2; BrCH2COBr 598-21-0; BrCH2CO2Et Abstr. 1954,48, 3930g (30) Wollenberg R H.;Albizati K. F.; peries R J. am Chem. Soc. 1977,99,7365. (33) Heller. R b; Shrewsbury, M. A. J. biol Chem. 1976, 251, (31) Leusink. A. J.; Budding, H. A Marsman, J. W.J. Organomet. 3815. Chem. 1967,9,286. leusink A. J.; Budding, H. A; drenth (34) Kleinsek D. b, ranganathan S.; porter J. W. proc. Natl W. Ibid. 1968, 11, 541 Acad. sci. U.S.A 1977,74. 1431. (32) horgan S. W.; morgan D. D; Orchin M. J. Org Chem. 1973, (36) Shefer. S.; hauser sIapar, V.; Mosbach, E, W. J. Lipid Rea. 38,3801. 1972,13. 402, Tetrahedron Letters, Vol.28,No.13, pp 1385-1388,1987 0040-4039/87 $3.00 .00 Printed in Great Britain Pergamon Journals Ltd.
method B is The triflate 1 1 ot catalyst 3 72 % by 11 can hmpa ( 2 SYNTHESIS OF AN HMG-COA REDUCTASE INHIBITOR; in a one-pot A DIASTEREOSELECTIVEALDOL APPROACH ransformation J, E. Lynch*, R. P. Volante, R. V. Wattley, I. Shinkai 239 and Merck Sharp and dohme Research Laboratories P.O. Box 2000, Rahway, New Jersey 07065
according ' by heating to ABSTRACT and The synthesis of the 6-hydroxy- -lactone moiety of an HMG-CoA reductase smoothly in 90 inhibitor in its correct absolute configuration (94% ee) has been accomplished via a diastereoselective aldol reaction between aldehyde 4 and the magnesium detalls of the enolate of Claisen condensation. and hydroxyl directed reduction of the resulting -hydroxy-6-keto ester.
We would The discovery of mevinolin la1 and compactin 1b2 as potent inhibitors of 3-hydroxy- 3-methylglutaryl coenzyme A reductase has resulted in an intensive effort to develop 3 4 efficient laboratory syntheses of these and other related hypocholesterolemic agents. The key structural feature common to these HMG-CoA inhibitors Is the 9-hydroxy-&-lactone moiety Furthermore, it appears that this unit in its correct absolute configuration EnGI., 1986, 6 (that which corresponds to la and 1b is essential for enzyme inhibition Thus, we ,
, Y. B Yang.
v.; la R=CH3 1b R=H
as well as others7 ,have been interested in the development of an efficient method for for the synthesis of such a structural unit angeri. A,; Prior syntheses of 8-hydroxyl-& -lactone HMG-CoA reductase inhibitors have been accomplished by initial generation of an appropriate, non-racemic lactone "synthon" 385. 47, followed by the coupling of this unit to the remaining portion of the molecule at a Site removed from the asymmetric centers.6 these routes have typically suffered from either 5, 26, 1699. Inefficient generation of the asymmetric synthon and/or inefficient coupling reactions
1385 1386
Our strategy is based on the use of a diastereoselective aldol reaction followed by a acknowledge claisen condensation for the generation of the 5(s) stereocenter of keto-alcohol from 3 ments and aldehyde 4 The 5(S)hydroxyl function of 3 would then be expected to direct the reduc- manuscript tion of the 3-keto group to the desired 3(R) stereochemistry of 2.
1, Albert J.; L Staplr Liesct
2. Endo,
Y. Y. 3. Compa 1983, 2 3 4 A long standing problem in the preparation of simple (a-unsubstituted B-hydroxy N.L. esters (adds) has been the lack of aldehyde facial selectivity in the aldol reaction with chiral acetate enolates However several solutions to this problem have recently 4. Lee, appeared.' We wish to report here the use of one of these methods for the diastereo C.H.; s.; I selective synthesis of the biphenyl HMG-CoA inhibitor 2.10 1985, The magnesium IIenolate of S(+)-2-acetoxy-l,1.2-triphenylethanol (5) (1.1 equiv) 5. stokk was condensed with aldehyde In tetrahydrofuran at -78° via the procedure of Braun and A.A. Devant 9 to produce the diastereomers in 93% yield (SS:SR=97:3, as determined by PMR willa and HPLC transesterification of 6 with 1.05 equivalent of sodium methoxide analysis.13 6. sletz in methanol gave methyl ester 7 (S/R 97:3) In 95% yield Treatment of ester 7with E.G.; 3 equivalents of lithio-t-butyl acetate tetrahydrofuran at -30 to 40°C for 1 hr. in 7. Prugh resulted in the formation of 6-keto-&-hydroxy ester 3 in 90% yield? Highly stereo- J.R. chem. specific reduction of the 6-keto functionality was effected using sodium 2435 borohydride-triethylborane in tetrahydrofuran-methanol (4:1) at -78° giving diol 8 in 1984 Lett. 93% yield. 6,11,12 The diol ester was saponified with sodium hydroxide in aqueous Tetra methanol, acidified to pH 3.8. and the resulting acid 9was lactonized by heating In 8. at for 8 afford in 85% yield determined toluene 90° hr to lactone 2 (3R55:3S5R 97:3 as Washington by HPLC analysis of the corresponding R(+)-a-methylbenzylamine derivatives).13 The ed. J Ne1 so enantiomeric purity of 2 could be enhanced to >99% ee by recrystallization of the C.H., R(+)-a-methylbenzylamine salt of the acid 9 prior to lactonization. 14 New Y 9. iwasa Tetra R.; Hagin E.;
10. G.E. Novel 79. 1387
I by a 3 from Acknowledgement: We wish to thank Hr. R. Reamer and ms. L. dimichele for NMR measure- ments and Mr. Ryan and Hr. E. Corley for chemical assistance, and Ms. Lee Dolan for e reduc- I K. ! manuscript preparation I I 1. Alberts, A.O.W., Chen, J.; Kuron, G Hunt, V.; Huff, J.; Hoffman C.; Rothrock, I J.; Lopez. H.; Joshua, H.: Harris, E.; Patchett, A.; monaghan, R.; Currie, S.; Stapley, E.; Albers-Schonberg, G Hensens, 0.; Hirshfield, J.: Hoogsteen, K.; Liesch, J.; Springer, J. Proc. Natl. Acad. Sci. USA 1980, 77 3957. i i 2. Endo, A.; Kuroda, H.; Tsujita, Y.; J. Antiblot, 1976, 29 1346. ! 3. Compactin-Hsu, C.-T.; Wang, N.-Y.; Latimer L.H.; Sih, C.J. J. Am. Chem SOC., ! 1983, 105, 593. Hirama M.; Uei M. ibid, 1982, 104, 4251. Girota, N.N.; Wendler, -hydroxy N.L. letrahedron Lett. 1982, 5501. girota N.N.Wendler, N.L. ibid 1983, 3687. Grieco, P.A.; Zelle R.E. lis R.; Finn, J. J. Am. Chem. soc. 1983 105 1403. reaction Mevinolin-Hirama H.; Iwashita, H. Tetrahedron lett.1983 recently 4. Lee, T.-J,; Holtz, W.J. Smith, R.L. J 0 chem 1982, 47 4750. KUO, C.H.; Patchett, A.A.: Wendler, N.L. ibid Yang, Y.L. Manna, S Falck, J.R., Yang, Y.-L. Tetrahedron Lett. 1984 3563; T.-J,; Lee, ibid 1985, 4995. I equiv) and 5. Stokker, G.E. Hoffman, W.F. Alberts. A.W.; cargoe Jr., E.J.; Deana, A.A.; Huff, J.W.; Novello, F.C.; Prugh, J.D.; Smith, R.L.; by PMR Gilfillan J.L. Willard, A.K., J. Med. Chem. 1985, 28, 347. 6. Sletzinger, H.; Verhoeven, T.R. Volante, R.P.; HcNamara, Corley, 7 N.M.; with E.G.; liu T.W.H., Tetrahedron Lett. 1985, 2951. 1 hr. stereo- 7. Prugh, J.D.; Deana, A.A., Tetrahedron Lett. 1982, 281. Yang, Y.-L. Falck, J.R. ibid 1982, 4305. Danishefsky, S Kerwin, J.R.; Kobayshi S. J. Amer, sodium Chem. soc. 1982, 104, 358. Prasad U.; Repic 0. Tetrahedron lett.1984 in 2435and 3391 rosen T.; Taschner, M.J.; Heathcock, C.H. J. Ora. Chem. 8 1984, 49, 3994. Majewski, M.; Clive, D.L.J.; Anderson, P.C., letrahedron aqueous Lett. 1984 2106. Guindon Y.; Yoakim, C.; Bernstein M.A.; Morton, H.E. in Tetrahedron Lett. 1985, 1185. 8. Morrison, J.D. and Mosher, H.S. "Asymmetric Organic Reactions" Am Chem. SOC. Washington D.C. 1976, p. 142-152. Soladle, G in 'Asymnetric Synthesis: Vol. 2. The ed. J.D. Morrison. 1983. Academic Press. New York N.Y.. p-188-199. Evans. D.A.; of the Nelson, J.V.; Taber,T.R. in Topics in Stereochemistry, 1982, 13, 1. Heathcock; C.H., in 'Asmtric Synthesis" Vol. 3 ed. J.D. Morrison. 1984 Academic Press; New York N.Y., pp 174-190.
9. lwasawa. N.: Mukaiyama.-- T. Chem. Lett.. 1983. 297. Braun. H.: Devant. Tetrahedron -Lett. 1984, 5031. Nagao, Y Yamada, S,; Kumagai T Ochiai, h.; Fujita,E. J. Chem. SOC. Chem. comm 20 1418 (1984). Nagao, V.; Hagiwara, Y.; Kumagai, T.; Ochiai, R.; Inoue, T.; Hashimto, K.; Fujita, knopfel Chem.; 51, 2393 (1986). Helmchen, 6.; Leifkauf, U.; Tauter Angew Chem. Int.Ed. Ingl.; 24 874 (1985).
I 10. G.E. Stokker, A.W. Alberts, P.S. Anderson, E.J. Cragoe, Jr., A.A. Deana, J.L. Gilfillan J. Hirshfield, W.J. Holtz W.F. Hoffman, J.W. Huff, T.J Lee, F.C. Novello, J.D.Prugh C.S. Rooney, R.L. Smith, and A.K. Willard, J.Med.Chem. 1986, 29, 170.
4 Tetrahedron LI 1388 I Printed in Grr
11. Narasaka, K.; Pal, F.C.; Tetrahedron, 1984, 2233. 12. The ratio of diol to diol was 98:2. 13. Enantiomeric and diastereomeric purity of 9 was obtained by conversion to the NOL corresponding R(+)-a-mcthylbcntylarnidcr 10 equivalents of 99.9 pure R(+)=a-mtby benzylamine at 90° for 12 hr) and HPLC separation on Dupont Zorbax silica eluting with hexane:diethylether:methyl-t-butylethtr: acetic acid 4:3:6:1 14. The stereochemical integrity of methyl ester 7 was determined by proton magnetic resonance In the presence of the chiral solvating agent L-phenyl tri fluoromethylcarbinol and/or Eu HFC)3. 15. Although the Claisen condensation could be performed directly on 6' giving 3 in 85% yield, the higher temperature required for this reaction 25°C resulted In epimerization of the newly generated chiral center. 16. Satisfactory infrared and proton magnetic resonance spectra were obtained for all compounds. CHFN analysis was obtained for compounds 2, 10 and 9.
(Received in USA 26 December 1986)
La synthes l'action de 1'0 1-5 dicarbonyles 11- 12 imidates . Dans ce tr par condensatic Les result reactions 2 t qliatre produitr nes-1,5 15 4 5 benzimidazoles Cependant pendant quatre excedant 60% e La determined de leurs carac references 15- 1
prioritpar d Soit par une r dans les condi et le xylene sition thermic