AU667442B2 - A pyrrolidylthiocarbapenem derivative - Google Patents

Description

i k 667442

AUSTRALIA

Patents Act 1990 COMPLETE SPECIFICATION STAN~DARD PATENT 4k .*a 0S 0 6 0...9 0 a 0 AppliCant(s): SHIONOGI SEIYAIKU KABUSHIKI KAISHA Invention Title: A PYRROLIDYLTHIOCARBAPENEM DERIVATIVE The following statement is a full description of this invention, including the best method of performing it known to me/us:

M.

BACKGROUND OF THE INVZNTION 1. Field of the Invention: The present invention relates to a new pyrrolidylthiocarbapenem derivative having a wide range of antibacterial spectrum, an antibacterial agent comprising the carbapenem derivative, a new pyrrolidine derivative which is an intermediate for producing the carbapenem derivative, and a method for producing the pyrrolidylthiocarbapenem derivative and the pyrrolidine derivative.

:09oa 2. Description of the Prior Art: Various compounds are known as carbapenems, a 15 kind of a p-lactam antibiotic. For example, imipenem, .meropenem, the mesylate (mesylamino), and the urea derivatives of a carbapenem as shown below are known.

:OH imipenem OH Me meropenem a -/Hs/\NHCH=NH

-M

2 OOH coOH mesylate urea OH Me derivative O e derivative OH Me OH leC NHS0 2Me 1-C_>-^NHCONH2 COOH

COOH

All of these compounds have a wide range of antibacterial spectrum, and are effective against both Gram-positive bacteria and Gram-negative bacteria. A 2 carbapenem derivative having a wider range of antibacterial spectrum and a stronger antimicrobial activity has been desired.

SUMMARY OF THE INVENTION The pyrrolidylthiocarbapenem derivative of this invention is represented by Formula I: OXI R1 R 4 1 R 2

NSO

2

N<R

3 :a OOX 2 I 0Jx 2

(I)

wherein R 1 is hydrogen or lower alkyl; R 2

R

3 and R 4 are hydrogen, lower alkyl which can be substituted or an amino protecting group independently, or R 2 and R 3 together with a nitrogen atom to which R 2 and R 3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or R 3 and R 4 together with .two 15 nitrogen atoms and one sulfur atom in the sasiegroup form a saturated or unsaturated cyclic group; i each cyclic group can further include at least one atom selected from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted;

X

1 is hydrogen or a hydroxy protecting gioup; X 2 is hydrogen, a carboxy protecting group, an ammonio group, an alkali metal or an alkaline-earth metal; and Y2 is hydrogen or an amino protecting group.

In another aspect of the present invention, the pyrrolidine derivative of the present invention is -3represented by Formula II: R4 *I R 2 yl -NSO

<R

3 N Y 2 II wherein R 2

R

3 and R 4 are hydrogen, lower alkyl which can be substituted, or an amino protecting group independently, or R 2 and R 3 together with a 5 nitrogen atom to which R 2 and R 3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or

R

3 and R 4 together with two nitrogen atoms and one sulfur atom in the sufamide group form a saturated or unsaturated cyclic group; each cyclic group can further include at least one atom selected from the group Sconsisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; Y 1 is hydrogen or a mercapto protecting group; and Y2 is hydrogen or an amino protecting group.

Alternatively, the present invention provides a method for producing a pyrrolidine derivative represented by Formula II: R4 1 2 yuS NSO 2 N< 3

NY

2

(II

4 wherein R 2

R

3 and R 4 are hydrogen, lower alkyl which can be substituted, or an amino protecting group independently, or R2 and R 3 together with a nitrogen atom to which R2 and R 3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or

R

3 and R4 together with two nitrogen atoms and one sulfur atom in the sufamide group form a saturated or unsaturated cyclic group; each cyclic group can further include at least one atom selected from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; yl is hydrogen or a mercapto protecting group; and Y2 is hydrogen or an amino protecting group; and 4 15 the method comprises the steps of: converting a o" a hydroxy group at the 4-position of a 4-hydroxypyrrolidine-2-carboxylic acid derivative into a mercapto group; converting a carboxy group at the 2-position into a hydroxymethyl group; converting a hydroxy group te in the hydroxymethyl group into an amino group or a sulfamoyl group; and converting the amino group into a sultamoyl group.

.Alternatively the present invention provides 25 a method for producing a pyrrolidylthiocarbapenem c derivative comprising the step of: allowing a carbapenem derivative to react with the pyrrolidine derivative of Formula II to obtain the pyrrolidylthiocarbapenem derivative of Formula I; the carbapenem derivative being represented by Formula III: I I 1 OX' R' x

M

coox 2 wherein R1 is hydrogen or lower alkyl; X 1 is hydrogen or a hydroxy protecting group; X 2 is hydrogen, a carboxy protecting group, an ammonio group, an alkali metal or an alkaline-earth metal; and X 3 is a leaving 5 group reactive ester group of hydroxy, alkylsulfinyl arylsulfinyl, alkylsilfonyl, or arylsulfonyl).

*4 *q

COC

Thus, the invention described herein makes possible the advantages of providing a new carbapenem derivative having a strong antimicrobial activity and a wide range of antibacterial spectrum, and a method for producing the carbapenem derivative; providing a new pyrrolidine derivative which is an intermediate for producing the carbapenem derivative, so*. 15 and a method for producing the pyrrolidine derivative; and providing an antibacterial agent comprising the carbapenem derivative.

These and other advantages of the present invention will become apparent to those skilled in the art upon reading and understanding the following detailed description.

6 DESCRIPTION OF THE PREFERRED EMBODIMENTS Followings are abbreviations used herein: Ac acetyl Alz all'loxycarbonyl Boc t-butoxycarbonyl Et ethyl Ft phthalyl Me methyl Ms methanesulfonyl NPrc protected amino Ph phenyl PMB p-methoxybenzyl Pmz p-methoxybenzyloxycarbonyl :t .PNB p-nitrobenzyl Pnz p-nitrobenzyloxycarbonyl Tr trityl Ts p-toluenesulfonyl A preferred scope of each group herein is as S. follows: The number of carbon atoms of "lower alkyl" 25 is 1 to 6. Examples of such an alkyl group include methyl, ethyl, n-propyl, iso-propyl, n-butyl, t-butyl, pentyl and hexyl. The number of carbon atoms of the lower alkyl is preferably 1 to 4. The most preferred lower alkyl is methyl or ethyl. Examples of a substituent of "a substituted lower alkyl" include hydroxy, alkoxy, amino, acylamino, lower alkylamino, carbamoyl, lower alkylcarbamoyl, carbamoyloxy, lower alkylcarbamoyloxy and cyano. The number of carbon atoms of r 7 "aralkyl" is 7 to 15. Examples of "an amino protecting group" and "a hydroxy protecting group" include lower alkoxycarbonyl, lower alkenyloxycarbonyl, halogenoalkoxycarbonyl, aralkyloxycarbonyl, trialkylsilyl and diazo. An example of the lower alkoxycarbonyl includes t-butyloxycarbonyl; an example of the lower alkenyloxycarbonyl includes allyloxycarbonyl; examples of the halogenoalkoxycarbonyl include 2 -iodoethyloxycarbonyl and 2 ,2,2-trichloroethyloxycarbonyl; examples of the aralkyloxycarbonyl include benzyloxycarbonyl, p-methoxybenzyloxycarbonyl, o-nitrobenzyloxycarbonyl, p-nitrobenzyloxycarbonyl and diphenylmethoxycarbonyl; examples of the trialkylsilyl include trimethylsilyl, triethylsilyl and t-butyldimethylsilyl.

In a definition of a group represented as follows: S4 /R NS02N, N

R

3 a saturated or unsaturated cyclic group formed from R 2 and R 3 together with a nitrogen atom to which R 2 and R 3 are bonded can be, a saturated or unsaturated 3 to 8 membered residue further having one or more of nitrogen, sulfur and/or oxygen atoms, if necessary, and a or 6 membered monocyclic residue including a hetero atom is preferable. The examples include pyrrolidin-1-yl, pyrrol-1-yl, imidazolidin-l-yl, imidazol-l-yl, pyrazolidin-l-yl, pyrazol-l-yl, piperidino, dihydro- or tetrahydropyridin-1-yl, piperazino, pipera- 8 zin-l-yl which may have a substituent at the 4position, morpholino anc thiomorpholino. Th groups may be substituted for one or more, preferably one or two, of the following groups: amino, protected amino, carbamoyl, lower alkyl, hydroxy, protected hydroxyl, lower alkoxy, oxo, lower alkylsulfonyl, hydroxy lower alkyl, carbamoyl lower alkyl, lower alkoxycarbonyl and cyano. Moreover, when the cyclic group it iiuidazolidin-l-yl, pyrazolidin-l-yl or piperazin-l- thi imino moiety thereof may be protected by a imii t ,,uecting group which is known in the art.

In the definition of the group IIa, a saturated or unsaturated cyclic group formed from R and 15 R 4 or R 3 and R 4 can be a saturated or unsaturated 5 to 7 membered residue having 2 to 3 nitrogen atoms and one sulfur atom and if necessary, having an another hetero atom such as an oxygen atom, and 5 to 6 membered monocyclic residue including a hetero jtom is preferable.

.4 5 Such a residue may include, if necessary, a substituent such as lower alkyl, halogen, lower alkoxy, acyloxy, hydroxy, amino, lower alkylamino, acylamino and oxo, and/or an unsaturated bond. The examples include 1,1- 4 dioxothiadiazinyl, 1,1-dioxodihydrothiadiazinyl, 25 1,1,3-trioxodihydrothiadiazinyl, 1,1-dioxothiadiazolizinyl, 1,1-dioxothiadiazolinyl, and 1,1,3-trioxothiadiazolinyl.

The "carboxy protecting group" is selected from those used in the art and serve the function of blocking the carboxyl group while reactions are carried out at other sites of the molecule. Such group generally contains less than about 19 carbon atoms and bind i.

9to a carboxyl group reversibly without affecting the other parts of the molecule. Typical examples include following groups: optionally substituted C1 C 8 alkyl, for example, methyl, methoxymethyl, ethyl, ethoxymethyl, iodomethyl, propyl, isopropyl, butyl, isobutyl, ethoxyethyl, methylthioethyl, methanesulfonylethyl, trichloroethyl, t-butyl, and the like; optionally substituted C 3

C

8 alkenyl, for example, propenyl, allyl, isoprenyl, hexenyl, phenylpropenyl, dimethylhexenyl, and the like; optionally substituted

C

7 C19 aralkyl, for example, benzyl, methylbenzyl, dimethylbenzyl, methoxybenzyl, ethoxybenzyl, nitrobenzyl, aminobenzyl, diphenylmethyl, phenylethyl, trityl, di-t-butylhydroxybenzyl, phthalidyl, phenacyl, S. 15 and the like; optionally substituted C6 C12 aryl, for example, phenyl, toluyl diisopropylphenyl, xylyl, trichlorphenyl, pentachlorophenyl, indaiLyl, and the like; optionally substituted C 1 C12 amino which is, an ester with acetone oxime, acetophenone oxime, acetoaldoxime, N-hydroxysuccineimide, N-hydroxyphthalimide, or the like; optionally substituted C 3 C12 hydrocarbonated silyl, for example, trimethylsilyl, dimethylmethoxysilyl, t-butyldimethylsilyl, and the like; optionally substitutel C3 C12 hydrocarbonated 25 stannyl, for example, trimethylstannyl, and the like.

Another carboxy protecting group is a pharmaceutically active ester forming group. Examples of such a group include following groups: 1-(oxgen-substituted)-C 2 to

C

15 alkyl groups, for example, a straight, branched, ringed, or partially ringed alkanoyloxyalkyl, such as acetoxymethyl, acetoxyethyl, propionyloxymethyl, pivaloyloxymethyl, pivaloyloxyethyl,, cyclohexaneacetoxyethyl, cyclohexanecarbonyloxycyclohexylmethyl, and the

I

10 like; C 3

C

15 alkoxycarbonyloxyalkyl such as ethoxycabonyloxyethly, and the like; C 2

C

8 alkoxyalkyl, such as methoxymethyl, methoxyethyl, and the like; C 4

C

8 2-oxacycloalkyls, such as tetraphdropyranyl, tetrahydrofuranyl, and the like; substituted C 8

C

12 aralkyls, for example, phenacyl, phthalidyl, and the like;

C

6

-C

12 aryl, for example, phenyl, xylyl, indanyl, and the like; C 2

C

12 alkenyl, for example, allyl, isoprenyl, 2-oxo-l,3-dioxolyl-4-yl-methyl, and the like.

Among the above, a protecting group used to block the carboxyl group during reactions is usually removed at the final step of the reaction, and therefore its structure is not essential. Thus, as one of skilled in the art can easily appreciate, the carboxy protecting 15 group can be selected from various equivalent groups including amides, acid anhydrides formed with carbonic acid or carboxylic acids, and the like as long as an aimed carboxyl group is protected properly.

An example of the lower alkyl includes tbutyl; examples of the lower alkenyl include allyl, isopentenyl and 2-butenyl; examples of the halogeno lower alkyl include 2-iodoethyl and 2,2,2-trichloroethyl; examples of the lower alkoxymethyl include methoxy- 25 methyl, ethoxymethyl and isobutoxymethyl; examples of the lower aliphatic acyloxymethyl include acetoxymethyl, propionyloxymethyl, butyryloxymethyl and pivaloyloxymethyl; examples of the 1-lower alkoxycarbonyloxyethyl include l-methoxycarbonyloxyethyl and 1-ethoxycarbonyloxyethyl; and examples of the aralkyl include benzyl, p-methoxybenzyl, o-nitrobenzyl, p-nitrobenzyl and diphenylmethyl. Examples of "an alkali metal' include lithium, sodium and potassium, and sodium or ar II~ II~ I~ IIII~ I I 11 potassium is preferred. Examples of "an alkaline-earth metal" include magnesium and calciur.i.

As "a mercapto protecting group", a conve.ntional one, acyl and aryl substituted lower alkyl such as benzyl, phenethyl, trityl and benzhydryl are included. As "a reactive ester group of hydroxy", a conventional one, a residue such as substituted or unsubstituted arylsulfonyloxy, lower alkanesulfonyloxy, halogeno lower alkanesulfonyloxy, dialkylphosphonyloxy, diarylphosphoryloxy and halogeno are included.

Ex:amples of the arylsulfonyloxy include benzenesulfonnyloxy, p-toluenesulfonyloxy, p-nitrobenzenesulfonyloxy and p-bromobenzenesulfonyloxy; examples of the lower S 15 alkanesulfonyloxy include methanesulfonyloxy and ethanesulfonyloxy; an example of the halogeno lower alkanesulfonyloxy includes trifluoromethanesulfonyloxy; an example of the dialkylphospheoryloxy includes diethylphosphoryloxy; an example of the diarylphosphoryloxy includes diphenylphosphoryloxy; and examples of the halogeno include chloro, bromo and iodo.

*e e An example of "an alkylsulfinyl gzoup" int*cludes methylsulfinyl, and an example of "an arylsulfi- S 25 nyl group" includes phenylsulfinyl.

The pyrrolidylthiocarbapenem derivative of the present invention is represented by the following Formula I: 12 0X1 R 1

R

4 2 2 NY 2 CxoX 2 I wherein R1 is hydrogen or lower alkyl; R 2

R

3 and R 4 are hydrogen, lower alkyl which can be substituted or an amino protecting group independently, and preferably R 4 is hydrogen, or R 2 and R 3 together with a 5 nitrogen atom to which R2 and R3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or we. R 3 and R 4 together with two nitrogen atoms and one sulfur atom in the sufamide group form a saturated or unsaturated cyclic group and each cyclic group can 10 further include at least one atom selected from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; X 1 is hydrogen or a hyu. c-cy protecting group; X 2 is hydrogen, a carboxy protecting group, an ammonio group, an alkali metal or 15 an alkaline-earth metal; and Y2 is hydrogen or an amino protecting group.

When the above pyrrolidylthiocarbapenem derivative I has a free -OH, -COOH, amino group, imino group, or substituted amino group, the pyrrolidylthiocarbapenem also includes pharmaceutically acceptable salts thereof. The same is the case with an intermediate compound for synthesizing the pyrrolidylthiocarbapenem derivative such as the pyrrolidins derivative I I 13 represented by Formula II. Examples of the pharmaceutically acceptable salts include a salt with a base, a salt with an acid, a salt with a basic or acidic amino acid and an intermolecular or intramolecular quarternary salt. Examples of the salt with a base include alkali metal salts such as sodium salt and potassium salt; alkaline-earth metal salts such as calcium salt and magnesium salt; ammonium salt; and organic amine salts such as triethylamine salt, pyridine salt, picoline salt, ethanolamine salt, triethanolamine salt, dicyclohexylamine salt, N,N'-dibenzylethylenediamine salt and dibenzylamine salt. Examples of the salt with an acid include inorganic acid addition salts such as hydrochloride, hydrobromide, sulfuric acid salt and 15 phosphoric acid salt; and organic acid addition salts such as formic acid salt, acetic acid salt, trifluoroacetic acid salt, maleic acid salt, tartaric acid salt, methanesulfonic acid salt, benzenesulfonic acid salt and toluenesulfonic acid salt. Examples of the salt with an amino acid include a salt with arginine, aspartic acid or glutamic acid.

The pyrrolidylthiocarbapenem derivative (I) of the present invention can be produced in the steps 25 of: by using, for example, 4-hydroxypyrrolid ine-2carboxylic acid or the derivative thereof as a starting material, obtaining a pyrrolidine derivative II represented by the following formula: R4 I R 2 YlS- i NSO 2 N R s Y 2 II 14 wherein R 2

R

3 and R 4 are hydrogen, lower alkyl which can be substituted, or an amino protecting group independently, and R4 is preferably hydrogen, or

R

2 and R 3 together with a nitrogen atom to which R 2 and R3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or R 3 and R together with two nitrogen atoms and one sulfur atom in the sufamide group form a saturated or unsaturated cyclic group; each cyclic group can further include one atom selected from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; yl is hydrogen or a mercapto protecting group; and Y2 is hydrogen or an amino protecting group; and b 15 allowing the obtained pyrrolidine derivative II to react with a carbapenem derivative represented by the following Formula III: ox' R' 33

CX

X

T

coox 2

(III)

wherein R 1 is hydrogen or lower alkyl; X 1 is hydrogen or a hydroxy protecting group; X 2 is hydrogen, a carboxy protecting group, an ammonio group, an alkali metal or an alkaline-earth metal; and X 3 is leaving group a reactive ester of hydroxy, alkylsulfinyl, arylsulfinyl, alkylsulfonyl, or arylsulfonyl).

The present invention also includes a pyrrolidine derivative represented by the following Formula II: R4 2 R4 ylNSO 2 N< R 3 I R 2 wherein R 2

R

3 and R 4 are hydrogen, substituent lower alkyl which can be substituted, or an amino protecting group independently, and R 4 is preferably hydrogen, or R 2 and R 3 together with a nitrogen atom to which R 2 and R3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or R 3 and R 4 together b** with two nitrogen atoms and one sulfur atom in the Ssufamide group form a saturated or unsaturated cyclic group; each cyclic group can further include at least one atom selected from the group consisting of oxygen, .o sulfur and nitrogen, and each cyclic group can be substituted; yl is hydrogen or a mercapto protecting group; and Y is hydrogen or an amino protecting group.

15 The pyrrolidine derivative II is prepared according to the steps of converting a hydroxy group at the 4-position of a 4-hydroxypyrrolidine-2-carboxylic acid derivative into a mercapto group; converting a carboxy group at the 2-positior into a hydroxymethyl group; directly sulfamidating a hydroxy group in the hydroxymethyl group or sulfamoylating it after converting it into an amino group; and removing the protecting group Y 1 if necessary. The order of these steps can be properly changed.

16 Synthesis of pyrrolidine derivative II Pyrrolidine derivative II is synthesized, for example, in the following process, but is not limited to.

Route 1 "0-"CO v 00 R

H

2 2 2

('JI)

R

a as d for ou NSON K 3 y1 1 In the above scheme, R 2 R and R 4 are the same as defined for Formula I, and R 5 is a group for forming an ester together with a carboxy group such as lower alkyl. yl and Y 2 are the same as defined for Formulas I and II, but denote a mercapto protecting group and an amino protecting group, respectively, at the intermediate of the reaction route.

In this process for example, 4-hydroxypyrrolidine-2-carboxylic acid derivative IV is first provided. A mesyl group or the like is introduced to the hydroxy group at the 4-position of compound IV, and then a protected mercapto group such as a tritylthio 17 group is introduced to the 4-position. In this way, a compound V is obtained. Then, a compound VI is obtained by reducing a carboxylate group at the 2position. An azide group is introduced to the compound VI, and the azide group is converted to an amino group, or phthalimide is reacted with the compound VI, and the formed phthalyl group is removed from the compound VI, thereby introducing an amino group at a position of the hydroxy group of the compound VI.

Thus, a compound VII is obtained. A sulfamoyl group is then introduced to the compound VII to obtain a compound II.

s** Furthermore, the process of Route 1 can be 15 variously modified. For example, after introducing a protected mercapto group to the 4-posit of the *compound IV, a carboxylate group is reducec then a sulfamide group is introduced to obt.in the compound II. Alternatively, after reducing the com- 20 pound IV, a protected mercapto group and a sulfamide group are successively introduced to obtain the compound II.

*9 SRoute 2 o HO.. X 4 0. N H ZNS NPrc 2 (IV) 2 (vw) Y 2

(IX)

y X p.4 R.2 Y y2 y2 jjI-) a I 18 In the above scheme, R 2

R

3 and R 4 are the same as defined in Formula I, and R 5 is a group for forming ester together with a carboxy group such as lower alkyl. Y 1 and Y2 are the same as defined in Formulas I and II, but denote a mercapto protecting group and an amino protecting group, respectively, at the intermediate of the reaction route. X4 is a hydroxy protecting group.

In this process, for example, a mesyl group or the like (represented by X 4 is first introduced to cthe 4-position of the 4-hydroxypyrrolidine-2-carboxylic acid derivative IV, then a carboxylate group is reduced to a hydroxymethyl group as is in Route 1 to obtain a 15 compound VIII. Then, a protected amino group such as a phthalimide group is introduced to a position of a hydroxy group in the hydroxymethyl group. Thus a compound IX is obtained. In introducing the protected amino group, it is effective to introduce a leaving group to the hydroxy group of the compound VIII to increase the reactivity. Next, a mercapto group protected by thioacetate and the like (represented by Y 1

S)

is introduced to the 4-position (see a compound and removing the protection to obtain a compound XI. By o 25 introducing a sulfamoyl group to the compound XI, a compound II-1 (a compound II wherein the -SY' at the 2-position of the pyrrolidine ring is SH) is obtained.

Furthermore, the process of Route 2 can be variously modified. For example, by introducing a protected mercapto group to the 4-position of the compound VIII, further introducing a sulfamoyl group and removing the protection, the compound II-1 is obtained.

19 Route 3

Y

2 2 In the above scheme, R 2 R and R are the sameas defined in'Formula I. Y 2 is the same as defined in Formula I but denotes a protecting amino group at the intermediate of the reaction route.

In this method, chloroformate or the like is first allowed to react with 4-hydroxypyrrolidine-2carboxylic acid IV-1 having protected nitrogen in the Next, after converting a hydroxy group in the hydroxymethyl group into a reactive ester and introducing a protected amino group, a compound XIII is obtained by removing the protection. A sulfamoyl group is introduced to the compound XIII resulting in a compound XIV, then, a protected mercapto group is introduced to a position of the hydroxy group at the 4-position. A compound II-1 is obtained by removing the protection of the mercapto group.

I

20 Synthesis of a pyrrolidylthiocarbapenem derivative The protection of the 4-position of the pyrrolidine derivative is removed to obtain an SH compound, if necessary, then, the pyrrolidine derivative is allowed to react with a carbapenem derivative represented by the following Formula III to give a pyrrolidylthiocarbapenem derivative I of the present invention: OX' R' 3 10 hydrogen or a hydroxy protecting group; X2 is hydrogen, a carboxy protecting group, an ammonio group, an alkali metal or an alkaline-earth metal; X3 is a leaving group 0 arylsulfinyl, alkylsulfonyl, or arylsufonyl).

The protection is removed from the compound I if necessary to give a compound having free carboxy, Shydroxy and/or amin R is hydrogen or lower aly; X is 0 hydrogen or a hydroxy protectmprising g roup; X is hydrogen, a carbapenem derivaotecting group, an ammono group, an alkali metal or an alkaline-earth metal; X is a leavomprising group pyrrolidyl thiocarbapenem derivative (including pharma- Sc (e.g.,tically acceptable salts thereof) of the presenty arylsulfinyl, alkylsulfonyl, or arylsufonyl).

00o 15 invencihn is administered as an antibacterial agent.

if necessary to give a compound having free carboxy, An administration method is in oral administration or parenteral adinistraion; as injection (a formulation An antibacterial agent comprising the pyrrolidylthiocarbapenem derivative A pharmaceutical composition comprising the pyrrolidylthiocarbapenem derivative (including pharmaceutically acceptable salts thereof) of the present invention is administered as an antibacterial agent.

An administration method is in oral administration or parenteral administration; as injection (a formulation 21 in an ampoule or vial, a liquid, a suspension or the like for an intravenous injection, an intramuscular injection, a drip infusion, or subcutaneous injection), an external or local administration agent (an ear drop, a nasal drop, an ophthalmic solution, an ointment, an emulsion, a spray, a suppository and the like), and an oral preparation. Preferably, the composition is administered by injection, through skin or mucosa.

The pharmaceutical composition includes at least 0.01% by weight of the pyrrolidylthiocarbapenem derivative and further includes an appropriate excipient, auxiliary agent, stabilizer, wetting agent, emulsifier, and other additives depending upon the administration method. These additives must be pharmaceutically 15 and pharmacologically accet-able materials which do not inhibit the effect of the pyrrolidylthiocarbapenem derivative and which show no adverse effects on patients. For example, lactose, stearic acid, magnesium stearate, clay, sucrose, cornstarch, talc, gelatin, agar, pectin, peanut oil, olive oil, cacao butter, ethylene glycol, tartaric acid, citric acid and fumaric acid can be contained in the oral preparation. For parenteral administration, a solvent alcohol, a buffer, methyl oleate, water or the like), a buffer 25 solution, a dispersing agent, a dissolving auxiliary agent, a stabilizer methyl p-hydroxybenzoate, ethyl p-hydroxybenzoate, sorbic acid or the like), an absorbefacient (mono- or dioctanoate of glycerin), an antioxidant, a perfume, an analgetic, a dispersing agent, an adverse effect inhibitor, an action potentiator (an agent for regulating absorption and elimination, an inhibitor for enzyme decomposition, a P-lactamasE inhibitor, and other kinds of antimicrobial agents) and the like can be contained in the formulation.

A dose of the pyrrolidylthiocarbapanem derivative of the present invention depends upon the age of a patient, the type and the state of the disease and the kind of compounds to be used. Generally, daily dose ranges from 1 my/patient to about 4000 mg/patient, but more can be administered if necessary. For exampie, a dose of 1 mg (the external application) is administered 4 times a day, and a dose of 1000 mg S(intravenous injection) is administered 2 to 4 times a 0 day to treat an infection.

g* e 15 Characteristics of the pyrrolidylthiocarbapenem derivative The characteristics of the pyrrolidylthiocarbapenem derivative of the present invention as an antibacterial agent will now be described as compared with same known compounds.

a Antimicrobial activity: A minimum growth ;inhibitory concentration and an effect for preventing bacterial 25 infection of the pyrrolidylthiocarbapenem derivative of the present invention are compared with those of meropenem (Japanese Laid Open Patent Publication No. 60-233076) and imipenem (Japanese Laid Open Patent Publication No. 55-9090), respectively to find that the derivative of the present invention is superior to meropenem against Gram positive bacteria and superior to imipenem against Gram negative bacteria. The derivative of the present invention has an antibacterial i i -mp 23 potency against Pseudomonas aeruginosa, a kind of a Gram negative bacteria, equal to or twice as that of imipenem, meropenem and the mesylamino derivative of a carbapenem (Japanese Laid Open Patent Publication No. 63-179876). When compared with the urea derivative of a carbapenem (Japanese Laid Open Patent Publication No. 62-155279), the derivative has equal to or twice the antibacterial potency against Gram positive bacteria, twice the potency against the Gram negative bacte- IC ria and twice to eight times the potency against Pseudomonas aeruginosa.

0* Rabbit nephrotoxicity test: ~An administration of the derivative of the o. 15 present invention of 250 mg per 1 kg of the body weight of a rabbit reveals no toxicity. The same result is obtained by an administration of meropenem. When 150 mg/kg of imipenem is administered, medium renal toxicity is revealed. Sugar and protein are found in urine and a white microgranular change in the kidney is found.

Rate of decomposition by mouse renal dehydropeptidase 1: 25 The enzymatic decor position rate of the pyrrolidylthiocarbapenem derivative of the present invention by the action of renal dehydropeptidase I is 76% of that of imipenem, 40% of that of meropenem to show higher Stability.

Solubility in water: The solubility in water of the derivative of the present invention is 10% or more in a form of free Y I ~~i 24 acid, enabling an intravenous injection. In contrast, the solubility of imipenem and meropenem is about 2% and they cannot be administered except for a drip infusion.

Pharmacokinetics in vivo: When the derivative of the present invention is intravenously injected to a cynomolgus (10 mg/kg), the half-life is 1.1 hours, a recovery from urine is 62.2%, and an integrated value of a concentration in blood is 24.9 pg-hr/ml. The half-life is 1.44 times, the recovery from urine is 1.36 times and the integrated value of a concentration in blood (Area under the curve: AUC) is 1.44 times as much as those of merope- 15 nem. The half-life is 1.87 times, the recovery from too* urine is 1.93 times, and AUC is 1.87 times as much as those of imipenem.

9*C* When the derivative is intravenously injected into a mouse (20 mg/kg), the recovery from urine is 36.3%, and the integrated value of a concentration in blood is 12.1 pg.hr/ml. The recovery from urine is 2.18 times and AUC is 2.32 times as much as those of meropenem. The recovery from urine is 1.15 times and 25 AUC is 1.37 times as much as those of imipenem. The recovery from urine is 1.48 times as much as that of mesylate derivative of meropenem.

In this way, the present invention provides a new pyrrolidylthiocarbapenem derivative having a wide range of antibacterial spectrum and a strong antimicr*'bial activity against both Gram positive bacteria Lnd Gram negative bacteria, an antibacterial agent (compo- 1 I II I e 25 sition) comprising the carbapenem derivative, and a method for preparing the carbapenem derivative. Furthermore, a new pyrrolidin derivative as an intermediate for preparing the carbapenem derivative and a method for preparing the same are provided.

A minimum bacterial growth inhibitory concentration and an effect for preventing bacterial infection of the pyrrolidylthiocarbapenem derivative of the present invention are compared with those of meropenem and imipenem, respectively to find that the derivative of the present invention is superior to meropenem against Gram positive bacteria and superior to imipenem against Gram negative bacteria. The derivative of the 15 present invention has an antibacterial potency against V.4 Pseudomonas aeruginosa, a kind of a Gram negative bacterium, equal to or twice of that of imipenem, meropenem and the mesylamino derivative of a carbapenem. When compared with the urea derivative of a carbapenem, the derivative has an equal or twice the antibacterial potency against Gram positive bacteria, twice the potency against the Gram negative bacteria and twice to eight tim's the potency against Pseudomonas aeruginosa. The pyrrolidylthiocarbapenem deriva- 25 tive is less toxic to an organism than the conventional carbapenem derivatives. Since the derivative decomposes slowly in a body, the antimicrobial effect thereof lasts for a longer period of time. Moreover, sincethe derivative has a higher solubility in water than the conventional carbapenem derivatives, it can be applicable for injection.

26 Following Examples are given to show the present invention, but not to limit the scope therof.

Preparative Example 1 of a pyrrolidine derivative PMz Pmz N lie 3 PMz je *fl*

C

C. *C C C

C

C

C.

CCSG

@e.

C.

IC A

CC

C C

C.

C

eGGS i.

U

C.

Pmz N

M

1 PMz Tr'- NFt 8 Pnz Ir 1 6 TrS N N, N NH 1 9 11 Pmz j 10 N N11SO 2 NI~e 2

IPMZ

rS> TrS> lie KNAH.)zTh5O2kNO Pnz 13 Pmz j 12 N NHSO 2 NH~m

IPMZ

H Mie Prnz 27 Step 1. Preparation of an 0-mesyl compound To a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-hydroxypyrrolidine-2-carboxylic acid methyl ester (227.2 g: 0.735 mole) in dichloromethane (1.3 liter) stirring at -30*C, triethylamine (112.5 ml: 1.1 eq.) and methanesulfonyl chloride (56.8 ml: 1 eq.) are added. The mixture is stirred at the same temperature for 15 minutes. The reaction mixture is successively washed with dilute hydrochloric acid and water, dried over magnesium sulfate, and concentrated in vacuo to give (2S,4R)-l-p-methoxybenzyloxy-carbonyl-4-methanesulfonyloxypyrrolidine-2-carboxylic acid methyl ester (280.1 Yield: 98%.

NMR 6 (CDC1 3 ppm: 3.02, 3.04(2 x s, 3H), 3.56, 15 3.78(2 x s, 3H), 3.81(s, 3H), 4.98, 5.08(ABq, J=12Hz, 1H), 5.04, 5.12(ABq, J=12Hz, 1H).

IR (CHC1 3 cm- 1 1755, 1709, 1620.

Step 2. Preparation of a tritylthio compound 20 To a solution of triphenylmethylmercaptan S. (107..02 g: 1.5 eq.) in dimethylformamide (350 ml), an oil suspension containing 60% sodium hydride (13.42 g: 1.3 eq.) is added with stirring at 0°C. The mixture is stirred at room temperature for 1 hour. The reaction 25 mixture is mixed with a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-methanesulfonyloxypyrrolidine- 2-carboxylic acid methyl ester (100 g: 0.258 mole) in ml) with stirring at 0 0 C. The mixture is stirred at 60°C for 30 minutes. The reaction mixture is poured into cold dilute hydrochloric acid, and extracted with ethyl acetate.

The extract is successively washed with water and brine, dried, and concentrated in vacuo. The residue I 28 is purified by silica gel column chromatography (toluene :ethyl acetate =5 to give (2S,4S)- 1 -p-rethoxybenzyloxycarbonyl-4-tritylthiopyrrolidile- 2 carboxylic acid methyl ester (127.1 Yield: 87%.

NMIR 6 (CDCl 3 ppm: 3.50, 3.71(2 x s, 3H), 3.78, 3.84(2 x s, 3H), 4.87, 5.13(ABq, 3=12Hz, 1H), 4.89, 5.13(ABq, 3=12Hz, 1H).

IR V (CHCl 3 cm- 1 1750, 1700, 1618.

Step 3. Preparation of a methylol compound To a solution of (2S,4S)-l-p--methoxybenzy- :00000loxycarbonyl-4-tritylthiopyrrolidine-2-carboxylic acid methyl ester (127.1 g: 0.224 mole) in tetrahydrofuran liter), lithium borohydrLde 88 g: 1 eq.) is added .wo% 15 with stirring at room temperature. The mixture is stirred at 60*C for 30 minutes. The reaction mixture is allowed to cool to room temperature and water (100 ml) is a'dded in small portions with stirring. The formed precipitate is removed by filtration and the filtrate is concentrated in vacuo. The residue is dissolved in dichloromethane, dried over magnesium sulfate, and concentrated under reduced pressure. The residue is washed with ether to give (2S,4S)-1-p- 0 methoxybenzyloxycarbonyl- 4-tritylthiopyrrolidine-2methanol as white crystals (82.3 Yield: 68%.

NMR 6 (CDCl 3 ppm: 3.84(s, 3H), 4.93, 4,99(ABq, 3=12Hz, 2H).

IR Y (CHCl 3 cm- 1 3400, 1668, 1610.

Step 4. Preparation of a mesyl compound A solution of (2S,4S)-1-p-methoxybenzyloxycarbonyl-4-tritylthiopyrrolidinp -2-methanoI (22.33 g: 41.37 mmole) is diluted with dichloromethane (300 ml) 29 and th3 mixture is cooled to -30 0 C. To this mixture, triethylamine (6.92 ml: 1.2 eq.) and methanesulfonyl chloride (3.52 ml: 1.1 eq.) are added, and the mixture is stirred for 20 minutes. The reaction mixture is successively washed with dilute hydrochloric acid and water, dried over magnesium sulfate, and filtered. The filtrate is concentrated in vacuo to give crude 4S) -l-p-methoxybenzyloxycarbonyl-4-tritylthiopyrrolidine-2--methanol methanesulfonate (27.81 g: 45.02 mmole). Yield: 100%.

NMR 8 (CDCl 3 ppm: 2.89(s, 3H), 3.81, 3.83(2 x s, 3H), 4.85 to 5.07(m, 2H).

JIR y (CHC1 3 cm- 1 1725, 1690, 1610.

a eae 15 Step 5. Preparation of an azide compound assetsTo a solution of (2S,4S)-1-p-methoxybenzyloxycarbonyl- 4--tritylthiopyrrolidine- 2-methanol methanesulfonate (27.81 g) in dimethylformamide (120 ml), a solution (12 ml) of sodium azide (3.50 g: 53.8 mmole) in water is added. The mixture is stirred at 80 0 C for 0 s9 8 hours. The reaction mixture is poured into ice water 0.1and extracted with ethyl acetate. The extract is successively washed with water and brine and conc:,en- 00 2 trated. The residue is purified by silica gel column S0o* 25 chromatography to give (2S,4S)-lLp-methoxybenzyloxycarbonyl-2-azidomethyl-4-tritylthiopyrrolidine (17.27 g: 30.64 mmole). Total y-ield of Steps 4 and 5: 74%.

NMR 5 (CDCl 3 ppm: 3.84(s, 3H), 4.82 to 5.15(m, 2H).

IR y' (CHCl 3 cm'1: 2105, 1685.

Step 6. Preparation of an amino compound A solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl-2-aziiomethyl-4--tritylthiopyrroli.dine 30 (17.27 g: 30.64 mmole) in a mixture of ethyl acetate (150 ml), methanol (200 ml), and acetic acid (2.63 ml: 46 mmole) is subjected to conventional hydrogenation over 5% palladium on carbon (5 After the reaction, the catalyst is filtered off and the filtrate is concentrated in vacuo to give (2S,4S)-l-p-methoxybenzyloxycarbonyl- 2-aminomethyl-4-tritylthiopyrrolidine acetate (17.33 g) as a residue. The residue is dissolved in dichloromethane, washed with aqueous sodium hydrogen carbonate, and concentrated to give (2S, 4S )-2-aminomethyl-l-p-methoxybenzyloxycarbonly- 4 tritylthiopyrrolidine (16.82 g).

Step 7. Preparation of a phthalimido compound 15 Crude (2S, 4S)-l-p-methoxybenzyloxycarbonyl- 4 tritylthiopyrrolidine-2-methanol methanesulfonate (115.4 g) produced from (2S, 4S)-l-p-methoxybenzyloxycarbonyl-4-tritylthiopyrrolidine-2-methanol (96.24 g: :178 mmole) in the same manner as in the above-mentioned Step 4 is dissolved in dimethylformamide (1 liter).

0 After adding potassium phthalimide (65.94 g: 2 eq.), the mixture is stirred at 100 0 C for 1 hour. The reaction mixture is poured into ice water and extracted *00 *4 with ethyl acetate. The extract is successively washed with water and brine, and concentrated. The residue is purified by silica gel column chromatography (toluene ethyl acetate) to give (2S,4S)-l-p-methoxybenzyloxycarbonyl-2-phthalimidomfethyl4tritylthiopyrrolidine (99.4 Yield: 83.5%.

NMR 6 (CDCl 3 ppm: 3.78, 3.84(2 x 8, 3H), 4.65 to 5.00(m,. 2H).

IR Y (CHCl 3 cm- 1 1770, 1712, 1693, 1611.

31 Step 8. Removal of a phthalyl group To a solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl-2-phthalimidomethyl-4-tritylthiopyrrolidine (752 mg: 1.124 mmole) in a mixture of dichloromethane (3 ml) and methanol (12 ml), hydrazine hydrate (109 pl: 2 eq.) is added. The mixture is heated for hours. The reaction mixture is concentrated in vacuo. The residue is dissolved in dichloromethane ml) and the solid is filtered off. The filtrate is washed with water and concentrated in vacuo.

The residue is recrystallized from a mixture of dichloromethane and methanol to give (2S,4S)-l-pmethoxybenzyloxycarbonyl-2-aminomethyl-4-tritylthiopyrrolidine (471 mg). Yield: 78%. mp. 165 to 167*C.

15 NMR 6 (CDCl 3

:CD

3 0D=2:1) ppm: 3.46(s, 3H), 4.96, 4.89(ABq, J=12Hz, 2H).

IR Q (CHC1 3 cm- 1 1683, 1610.

Step 9. Preparation of a dimethylsulfamoyl compound A solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl-2-aminomethyl-4-tritylthiopyrrolidine (12.44 g: 23.13 mmole) in dichloromethane(70 ml) is cooled to -78°C. After adding triethylamine (4.21 ml: 1.3 eq.) and dimethylaminosulfonyl chloride (2.73 ml: 25 1.1 the mixture is warmed to room temperature over about 1 hour. The reaction mixture is successively washed with dilute hydrochloric acid and brine, and concentrated to give crude (2S,4S)-1-pmethoxybenzyloxycarbonyl-2-N,N-dimethylsulfamoylaminomethyl-4-tritylthiopyrrolidine (15.02 Yield: 100%.

1 32 Step 10. Preparation of a mercapto compound by deprotection To a solution of (2S,4S)-1-p-methoxybenzyloxycarbonyl-2-N,N-dimethylsulfamoylaminomethyl-4tritylthiopyrrolidine (3.55 g: 5.5 mmole) in a mixture of dichloromethane (70 ml) and methanol (35 ml), a solution of pyridine (0.66 ml: 1.5 eq.) and silver nitrate (1.40 g: 1.5 eq.) in water (3.5 ml) is added under ice cooling. The mixture is stirred for 10 minutes. The reaction mixture is poured into water and extracted with dichloromethane. The extract is dried over magnesium sulfate, bubbled with hydrogen *fl.

sulfide, and filtered to remove solid. The filtrate is concentrated in vacuo and the residue is purified by 15 silica gel column chromatography (toluene ethyl

S

eo acetate) to give (2S,4S)-l-p-methoxybenzyloxycarbonyl-2-N,N-dimethylsulfamoylaminomethyl-4-mercaptopyrrolidine (1.93 Yield: 87.0%.

0. NMR 6 (CDC13) ppm: 2.77(s, 6H), 3.81(s, 3H), 5.00 to 5.12(m, 2H).

IR Y (CHC13) cm- 1 3380, 1690, 1610.

Step 11. Preparation of a sulfamoyl compound .To a solution of chlorosulfonyl isocyanate 25 (3.95 ml: 45.4 mmole) in dichloromethane (70 p-methoxybenzyl alcohol (5.66 ml: 45.4 mmole) is added at -50 0 C. The mixture is stirred at -50°C for minutes. The resulting solution of p-methoxybenzyloxycarbonylsulfamoyl chloride is added to a solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl-2-aminomethyl- 4-tritylthiopyrrolidine (obtained in the above Steps 6 or 8) (12.21 g: 22.7 mmole) and triethylamine (6.38 ml: 45.6 mmole) in dichloromethane (300 ml) at -78°C, and i 33 the mixture is stirred for 10 minutes, successively washed with dilute hydrochloric acid and brine, and concentrated in vacuo. The residue is purified by silica gel column chromatography to give (2S,4S)-l-pmethoxybenzyloxycarbonyl-2-p-methoxybenzyloxycarbonylsulfamoylaminomethyl-4-tritylthiopyrrolidile (16.31 g).

Yield: 91.6%.

NMR '5 (CDCl 3 ppm: 3.78(s, 3H), 3.81, 3.83(2 x s, 3H), 4.98, 4.89(ABq, J=l2Hz, 2H), 5.09, 5.03(ABq, J=l2Hz, 2H).

IR 9 (CHCl 3 cm- 1 3390, 1740, 1685.

*fee Step 12. Preparation of a mercapto compound by deprotection 15 To a solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl2-p-methoxybenzyloxcycarbonylsulf amoylaminoinethyl-4-tritylthiopyrrolidine (2.35 g: 3.13 mmole) in a mixtureof dichloromethane (60 ml) and methanol ml), a solution of pyridine (0.38 ml: 4.75 mmole: 1.-5 eq. arid silver nitrate 80 g: 1. 5 eq. in water (2 ml) is added under ice cooling. The mixture is 5. stirred for 10 minutes. The reaction mixture is poured into water and extracted with dichloromethane. The extract is dried over magnesium sulfate and filtered.

*000: 25 Hydrogen sulfide is passed through the filtrate and the resulting precipitate is filtered off. The filtrate is concentrated in vacuo and the residue is purified by silica gel column chromatography to give (2S, 4S )--p-methoxybenzyloxycarbonyl-2-p-methoxybenzyloxycarbonylsulfamoylaminomethyl-4-mercap-topyrrolidine (1.56 Yield: 92.4%.

NMR 5 (ODC1 3 ppm: 2.42 to 2.58(m, lH), 3.80(s, 6H), 5.08, 5.02(ABq, J=12Hz, 2H), 5.12i 5.07(Asq, J=2GHz, 34 2H).

IR (CHC1 3 )cnf 1 3380, 1740, 1685, 1610.

Step 13. Pre2_atino nNmehlcmon To a solution of (2S,4S)-1-p.-methoxybenzyloxycarbonyl-4 -tritylthio-2- (p-methoxybenzyloxycarbonylaminosulfonylaminomethyl )pyrrolidine (2.06 g: 2.63 mmole) in direthylformamide (15 ml), a solution of iM-lithium bis(trimethylsilyl )amide in tetrahydrofuran (2.76 ml: 1.05 eq.) is added with stirring under ice cooling. After stirring for 1 hour, iodomethane (491 p1: 3 eq.) is added. The mixture is stirred at the same temperature for 3 hours. The reaction mixture #Gob poured into a mixture of ethyl acetate and aqueous sodium sulfite and the ethyl acetate layer is taken.

The organic layer is sucuessively washed with water and brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate 4:1) to give (28,4S)-l-p-methoxybenzyloxycarbonyl-4-tritylthio- 2 9 9.(N-p-methoxybenzyloxycarbonyl-N-methylaminosulfonlP anino)methylpyrro i dine (1.51 Yield: 72-..

MR (CDCl 3 ppm: 1.4 to 1.6(m, 1H), 1.9 to 2.1(m, 9 09 a 1H), 2.5 to 3.3(m, 4H), 3.23(s, 3H), 3.5 to 3.8(m, 1H), 00*0: 25 3.76(s,~ 3H), 3.81(s, 3H), 4.93(ABq,J=10.4Hz, 2H), J=15.2Hz, 2H), 6.35 to 6.55(m, 1H), 6.8 to 23H).

IR Y) (CHCl 3 cmnf1: 1727, 1695.

Step 14. Preparation of a mercapto compound by__deprotection To a solution of (2S,4S)-1-p-methoxybenzyloxycarbonyl -4-tritylthio-2- p-methoxybenzyloxycarbo- 35 nyl-N-methylaminosulfonyl)aminomethylpyrrolidine g: 1.88 mmole) in a mixture of dichloromethane (4 ml) and methanol (10 ml), a solution of pyridine (381 p 1: 2.5 eq.) and silver nitrate (640 mg: 2 eq.) in water (6 ml) is added with stirring under ice cooling. The mixture is stirred at the same temperature for 30 minutes. The reaction mixture is diluted with dichloromethane, washed with water, dried over magnesium sulfate, and concentrated in vacuo to about 5 ml.

The residue is dissolved in methanol (10 ml) and hydrogen sulfide is bubbled through it. The mixture freed goo* from solid by filtering is concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate 2:1) to give (2S,4S)-l-p- 15 methoxybenzyloxycarbonyl-4-mercapto-2-(N-p-methoxybenzyloxycarbonyl-N-methylaminosulfonyl)aminomethylpyrrolidine (866 mg). Yield: 83%.

NMR 6 (CDC13) ppm: 1.6 to 1.8(m, 1H), 2.3 to 2.6(m, 1H), 2.9 to 3.4(m, 5H), 3.3(s, 3H), 3.8(s, 6H), 20 3.8 to 4.2(m, 1H),-6.3 to 6.6(m, 1H), 6.88(d, J=8.6Hz, 2H), 7.2 to 7.4(m, 2H).

IR Y (CHC1 3 cm- 1 1690.

9 36 Peparative Example 2 of a pyrrolidine derivative HO, liO.

O

N 1 V Me 2 in3 PnzPnz Pnz TrS*.

N4

H

1 4 Pnz TrSPNz 1 Pnz Trs Pnz o

OC

C* C 0 0

C

6 T rS 2 7 Pnz nz Tr)-(,.NHSoc Pnz 8 9 N 0 2

NH

2 Pnz Step 1 Preparation of an 0-mesyl compound To a solution of (2S,4R)-l-p-nitrobenzyloxycarbonyl-4-hydroxypyrrolidine-2-carboxylic asid methyl ester (59.44 g: 0.183 mole) in dichioromethane (150 ml) stirring at -20CC, triethylamine (30.5 ml: 1.2 eq.) and methanesulfonyl chloride (17 ml: 1 eq. are added. The mixture is stirred at the same temperature for minutes. To the mixture is added ice water and ethyl acetate. The organic layer is taken, washed with water, dried over magnesium sulfate, and concentrated in vacuo to give (2S,4R)-l-p-nitobenzyloxycabonyl-4methanesulfonyloxypyrrolidine-2-carboxylic acid methyl ester (74.05 Yield: Quantitative.

-I i 37 NMR 6 (ODCd 3 PPM: 2.20 to 2.42(m, 1H), 2.55 to 3.80 to 4.05(m, 2H), 4.53(t, 3=7HZ, 1H), 5.06 to 5.40(m, 3H), 7.47(d, 3=9Hz, 1H), 7.51(d, 3=9Hz, 1H), 8.23(d, 3=9Hz, 2H).

IR (CHCl 3 cm- 1 1748, 1712, 1608.

Step 2. Preparation of a trityithio compound To a solution of tritylmercaptan (37.69 g: 1.5 eq.) in tetrahydrofaran (180 ml), an oil suspension containing 60% sodium hydride (4.73 g: 1.3 eq.) *is added with stirring at 0 0 C. The mixture is stirred at roow temperature overnight. A solution of (2S, 4R )-l-p-nitrobenzyloxycarbonyl-4-methanesulfonyl- 15 oxypyrrolidine-2-carboxylic acid methyl ester (36.58 g: 90.9 mmole) in tetrahydrofuran (180 ml) is added to the reaction mixture with stirring at 0 0 C, and the mixture is stirred at 600C for 30 minutes. The reaction mixture is poured into cold d5l~t~hydrochloric acid and extracted with ethyl acetate. The extract is successively washed with water and brine, dried, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate 9:1 to 4:1) to give (2S,4S)-l-p-nitobenzyloxycarbonyl- 4 tritylthiopyrrolidine-2-carboxylic acid methyl ester (25.48 Yield: 48.1%.

NMR 6 (CDC1 3 PPM: 1.63 to 2.35(m, 2H), 2.68 to 3.50(m, 3H), 3.60(s, 1.5H), 3.72(s, 1.5H), 4.02 to 4.15(m, 1H), 4.95 to 5.28(m, 2H), 7.10 to 7.52(m, 17H), 8.17(d, J=9Hz, 1H), 8.24(d, 3=9Hz, 1H).

IR (CHCl 3 cm- 1 1747, 1704, 1607.

38 Step 3. Preparation of a iethylol compound To a solution of (2S,4S)-l-p-nitrobenzyloxyr .rbonyl-4-tritylthiopyrrolidifle-2-carboxylic acid methyl ester (5 g: 9.01 mmole) in tetrahydrofuran (180 ml) stirring undr- ice cooling, a solution of ,-odium borohydride (2.3 g: 1.4 eq. in ethanol and a solution of lithium chloride 2. 76 g: 1 .5 eq. in tetrahydrofuran (60 ml) are added. The mixture is stirred at room temperature for 1 hour. The reaction mixture is poured into a mixture of ice water and ethyl acetate and extracted with ethyl acetate. The extract sucz,7essively washed with cold dilute hydrochloric acid, aqueous sodium hydrogen carbonate, and saturated brine, dried over magnesium sulfate, and concentrated 15 in vacuo. The residue is recrystallized from methanol to give (2S, 4S)-1-p-nitrobenzyloxycarbonyl-4-tri.tylthiopyrrolidine-2-methanol (15.9 Yield: 65.9%. mp.

122 to 1250C.

NMR i) (CDCl 3 ppm: 1.32 to 1.53(m, lR), 1.90 to 20 2.12(m, lH), 2.65 to 3.05(m, 3 3.32 to 3.84(m, 3H), 5.08, 5.17(ABq, J=l2Hz, 2H), 7.08 to 7.55(m, 17H), 8,26(d, J=9Hz, 2H).

y'(CHCl 3 cm- 1 31M0br, 1681, 1607.

Step 4. Preparation of a mesyl compound To a solution of (2S,4S)-1-p-nitrobenzyloxycarbonyl-4-tritylthiopyrrolidine-2-methanol (5.0 g: 9.01 mmole) in dichlaromethane (50 ml) stirring at triethylamine (1.63 ml: 1.3 eq.) and methanesulfonyl chloride (0.85 ml: 1.1 eq.) are added. The mixture is stirred at -15 to -10 0 C for 30 minutes. The reaction -mixture is poured into water and extracted with dichloromethane. The extract is successively 39 washed with dilute hydrochloric acid, aqueous sodium hydrogen carbonate, and water, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene: ethyl acetate to give (2S,4S)-l-p-nitrobenzyloxycarbonyl-4-trityltbt-opyrrolidine-2-methanol methanesuifonate (4.86 Yield: 85.2%.

NMR 6 (CDCl 3 PPM: 1.65 to 1.93(m, 1H), 2.00 to 2.26(m, lH), 2.68 to 2.92(m, 3H), 2.96(s, 3H), 3.78 to 3.98(m, 1H), 4.16 to 4.30(m, 1H), 4.38 to 4..52(m, 1H), s, 2H), 7.08 to 7.52(m, 17H-), 8.24(d, J=9Hz, 2H).

IR y (CHC1 3 1699, 1606.

3)c Step 5. Preparation of a phthalimido compound A solution of (2S,4S)-l-p-nitrobenzyloxycarbonyl-4-tritylthiopyrrolidine-2-methanol methanesulfonate (4.39 g: 6.93 mmole) and potassium phthalimide (2.57 g: 2 eq.) in dimethylformamide (30 ml) is stirred 20 at 70'C for 6 hours. The reaction mixture is poured *199*4into ice water and the precipitate is filtered off.

The precipitate is dissolved in ethyl acetate, washed with saturated. brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified byT 2 5 silica gel column chromatography (toluene ethyl acetate) to give (2S,4S )-l.-p-nitrobenzyloxycarbonyl-2phthalimi,3omethyl-4-tritylthiopyrrolidine 12 g).

Yield: 64.3*..

NMR 5 (CDCl 3 ppm: 1.40 to 2.30(m, 2H), 2.60 to 3.08(m, 2H), 3.10 to 3.40(m, 1H), 3.55 to 4.23(m, 3H), 4.92, 5.06(ABq, J=12Hz, 7.08 to 7.50 (in, 17H), 7.60 to 7.82(m, 4H), 8.10(d, J=9Hz, 1H), 8.19(d, J=9Hz, 1H).

40 IR Q (CHC1 3 cm- 1 1720, 1701, 1607.

Step 6. Removal of a phthalyl group To a solution of (2S,4S)-1-p-nitrobenzyloxycarbonyl-2-phthalimidomethyl-4-tritylthiopyrrolidine (10.46 g: 15.31 mmole) in a mixture of dichloromethane ml) and methanol (160 ml), hydrazine hydrate (1.53 ml: 2 eq.) is added, and the mixture is concentrated to remove dichloromethane by warming and refluxed for 3 hours and 15 minutes. The reaction mixture is concentrated in vacuo. The residue is diluted with dichloromethane and filtered to remove solid. The filtrate is washed with water, dried over magnesium sulfate, and concentrated in vacuo to give 15 crude (2S,4 ;)-l-p-nitrobenzyloxycarbonyl-2-aminomethyl- 4-tritylth.opyrrolidine (7.71 Yield: 91%.

NMR 6 (CDCl 3 :CDO3D=2:1) ppm: 1.46 to 3.76(m, 5.04, 5.12(ABq, J= 15Hz, 2H), 7.10 to 7.56(m, 17H), 8.12 to 8.30(m, 2H).

20 IR Q (CHC1 3 cm-1: 1695, 1606.

r Step 7. Preparation of an N-sulfamoy. compound A solution of (2S,4S)-1-p-nitrobenzyloxycarbonyl-2-aminomethyl-4-tritylthiopyrrolidine (4.7 g: 8.49 mmole) is dissolved in dichloromethane (45 ml) and cooled to a temperature of -70°C. To the mixture, a solution of diisopropylethylamine (3.4 ml: 2.3 eq.) and IM t-butoxycarbonylaminosulfonyl chloride (prepared from chlorosulfonyl isocyanate and t-butanol before hand) in dichloromethane (21 ml), and the mixture is stirred for 1 hour and diluted with ice water. The reaction mixture is successively washed with dilute hydrochloric acid and aqueous sodium hydrogen carbon- 41 ate, dried over magnesium sulfate, and concentrated.

The residue is purified by silica gel column chromatography (toluene :ethyl acetate) to give 2

S,

4 S)-l-pnitrobenzyloxycarbonyl-2-t-butoxycarbolyl-aminosulfonylaminomethyl-4-tritylthiopyrrolidine (1.49 g).

Yield: 24%.

NMR 6 (CDCJ.

3 ppm: 1.40 to 2.30(m, 2H), 1.44(s, 9H), 2.60 to 3.40(m, 5H), 3.71 to 3.95(m, lH), 5.08, 5.13(ABq, J=l2Hz, 2H), 6.27(br s, 1H), 7.07 to 7.55(m, 17H), 8.21(d, J=7Hz, l1H), 8.26(d, J.=7Hz, 1H).

Of (C 3 cm- 1 30 1737, 1695, 1606.

Step 8. Removal of a Boc group *41* To a solution of (2S, 4S)- l-p-nitrobenzyloxycarbonyl-2--t-butoxycarbonylaminosul fonylaminomethyl-4 tritylthiopyrrolidine (1.46 g: 2 mmole) in dichioromethane (5 ml) under ice cooling, anisole (2.4 ml) and trifluoroacetic acid (3.9 ml) are added. The mixture is stirred at room temperature for 2 hours. The reac- 20 tion mixture is diluted with ethyl acetate and ice water and extracted with ethyl acetate. The extract is successively washed with water and saturated brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is recrystallized from n-hexane to give (2S,4S)-l-p--nitrobenzyloxycarbonyl.-2-sulfamoylaminomethyl-4-tritylthiopyrrolidine (1.4 Yield: Nearly quantitative.

Nt-R 6 (OD1 3 ppm: 1'43 to 1. 70 1H) 2. 08 to 2.30(m, 1H), 2.65 to 3.50(m, 5H), 3.74 to~ 4.00(m, 1H), 5.03, 5.13(ABq, J=l5Hz, 21-f), 5.73(br s, 1H), 7.00 to 7.60(m, 17H-), 8.25(d, J=9Hz, 2H).

IR Q (CHC1 3 cm- 1: 3334br, 1688, 1607.

42 Step 9. Preparation of a mercapto compound by deprotection To a solution of (2S,4S)-l-p-nitrobenzyloxycarbonyl-2-sulfamoylaminomethyl-4-tritylthiopyrrolidine (668 mg: 0.95 mmole) in tetrahydrofuran (6 ml), a solution of pyridine (0.254 ml: 2.7 eq.) and silver nitrate (403 mg: 2.5 eq.) in water (2 ml) is added under ice cooling. The mixture is stirred at room temperature for 1 hour. The reaction mixture is diluted with dichloromethane (3 ml) and methanol (3 ml), and hydrogen sulfide is bubbled through it under ice cooling for 10 minutes. The resulting precipitate is removed by filtering. The filtrate is diluted with dichloromethane, washed with water, dried over magnesi- 15 um sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate) to give (2S,4S)-l-p-nitrobenzyloxycarbonyl-2-sulfamoylaminomethyl-4-mercaptopyrrolidine (233 mg). Yield: 63%.

20 NMR 6 (CDC13-CD30D) ppm: 1.42(t, J=7Hz, 1H), 1.65 to 1.93(m, 1H), 2.48 to 2 .70(m, 1H), 3.05 to 3.63(m, 4H), 3.93 to 4.16(m, 2H), 5.22(s, 2H), 7.53(d, J=8Hz, 2H), 8.23(d, J=8Hz, 2H).

IR (CHC1 3 cm- 1 3276br, 1692, 1607.

43 Preparative Example 3 of a pyrrolidine derivative MsO Ms0 MsO'..

Pnz Pnz Pnz t A K 2 6 Pnz Pnz Pnz NIS0W2NH2 Pn Step 1. Preparation of a methylol compound To a solition of (2S,4R)-1-p-nitrobenzyloxycarbonyl-4-methanesulfonyloxypyrrolidine-2-carboxylic acid methyl ester (79.4 g: 0.197 mmole) in a mixture of ethanol (300 ml) and tetrahydrofuran (150 ml), sodiui borohydride (10.44 g: 1.4 eq.) is added in small poro tions with stirring at 0 0 C. The mixture is stirred at S0 0 "C for 1.5 hours and at room temperature for 5 hours.

i To the reaction mixture under ice cooling, chloric acid (100 ml) is added, The mixture is diluted

S.

with water, and extracted with ethyl acetate. The extract is washed with brine, dried over sodium sulfate, and concentrated in vacuo. The residue is recrystallized from a mixture of dichloromethane and ether to give (2S,4R)-1-p-nitrobenzyloxycarbonyl-4methanesulfonyloxypyrrolidine-2-methanol (51.9 g).

Yield: NMR 6 (CDC13) ppm: 1.93 to 2.14(m, 1H), 2.32 to 2.48(m, 1H), 3.06(s, 3H), 3.53 to 4.28(m, 6H), 5.26(s, 2H), 7.53(d, J=9Hz, 2H), 8.24(d, J=9Hz, 2H).

44 IR (Cl-Id 3 cm-n 1 3404, 1698, 1607.

Step 2. Preparation of a tosy. compound To a solution of (2S,4R)-1-p-nitrobenzyloxycarbonyl-4-methanesulfonyloxypyrrolidine-2-methano.

(28.8 g: 77 mmole) in dichiloromethane (150 ml) under ice cooling, p-toluenesulfonyl chloride (19.11 g: 1.3 eq. triethylarnine (10.4 ml: 1.3 eq. and dimethylaminopyridine (0.94 g: 0.1 eq.) are added. The mixture is stirred at 25 0 C for 7 hours. The reaction mixture is diluted with ice water. The x"~sultant organic layer is taken, successively washed -ta aqueous sodium hydrogen carbonate and water, dre.% over magnesium sulfate, and concentrated in vacuo. The residue is recrystallized from n-hexane to give (2S, 4R )-l-p-nitrobenzyloxycarbonyl-4-methanesulfonyl- 0:0. oxypyrrolidine- 2-methanol p-toluenesulfonate (37.7 g).

Yield: 93%.

NMR 6 (CDC1 3 ppm: 2.20 to 2.50(m, 1H), 2.44(s, 3H), 20 3.05(s, 3H), 3.45 to 4.60(m, 5H), 5.18(s, 2H), 5.26(br 0.8. s, 1H), 7.34(d, J=8Hz, 2H), 7.50(d,J=8Hz, 2H), 7.75(d, J=8Hz, 2H), 8.23(d, .1=BHz, 2H).

.IR (CHCl 3 c 1 1700, 1599.

Step 3. Preparation of a phthalimido compound A mixture of (2S,4R)-l-p-nitrobenzyloxycarbonyl-4-methanesulfonyloxypyrrolidine-2-methanol ptoluenesulfonate (25 g: 47.3 mmole) and potassium phthalimide (17.52 g: 2 eq.) in dimethylformamide (250 ml) is stirred at 60*C for 7 hours. The reaction mixture is poured into ice water and filtrated. The resulting precipitate is dissolved in ethyl acetate, washed with saturated brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is recrystallized from methanol to give (2S,4R)-l-p-nitrobenzyJloxycarbonyl-2-phthalimidomethyl-4 -methanesulfonyloxypyrrolidine (18.76 Yield: 79%. mp. 121 to 123 0

C.

NMR (CDCl 3 ppm: 2.03 to 2.60(m, 2H), 3.02(s, 3H), 3.50 to 4.15(m, 4.40 to 4.63(m, 1H), 5.10, 5.29(ABg, J=l5Hz, 2H), 5.10 to 5.30(m, 1H), 7.46 (d, J=9Hz, 7,57(d, J=9Hz, 1H), 7.63 to 7.88(m, 4H), 8.20(d, J=9Hz, 2H-) IR y (CHCl 3 cm- 1 1773, 1715, 1605.

*'*Step 4. Preparation of an acetylthio compound solution of (2S,4R)-l-p-nitrobenzyloxycarbonyl-2-phthalimidomethyl-4-methanesulfonyloxypyrrolidine (10 g: 19.88 mmole) and potassium thioacetate (4.54 g: 2 eq.) in dimethylformamide (60 ml) is stirred at 60*C for 3 hours. The reaction mixture is pouted 0 into ice water (200 ml) and filtered. The precipitate W' 4, 20 is dissolved in ethyl acetate, dried over magnesium .*to sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate) to give (2S,4S)-l-p-nitrobenzyloxycarbony 1-2-ph thal imid omethyl -4-ace tylt hi opy rro lid in e (8.7 Yield: NMR (CDCl 3 ppm: 1.65 to 1.97(m, 1H), 2.47 to 2.67(m, 3.24 to 3.34(q, 1H), 3.73 to 4.24(m, 4H), 4.30 to 4.54(m, 1H), 5.02(dd, J=14Hz, >-7Hz, 1H), 5.20(d, J=14H4z, 1H), 7.42(d, J=9gHz, 1H), 7.45(d, J=9Hz, 1H), 7.60 to 8.86(m, 4H), 8.17(d, 3=9Hz, 2H).

IR (CHCl 3 cm- 1 1773, 1714, 1605.

46 Step 5. Removal of a phthalyl and an acetyl groups To a solution of (2S,4S)-l-p-nitrobenzyloxycarbonyl-2-phthalimidomethyl-4-acetylthiopyrrolidine (4.92 g: 10.18 mmole) in a mixture of dichloromethane (15 ml) and methanol (75 ml), hydrazine hydrate (1.53 ml: 3 eq.) is added. The mixture is warmed to remove dichloromethane and heated to reflux for 1 hour and 10 minutes. The reaction mixture is concentrated in vacuo. The residue is diluted with dichloromethane and filtered. The filtrate is washed with water, dried over magnesium sulfate, and concentrated in vacuo «to give crude (2S,4S)-l-p-nitrobenzyloxycarbonyl-2aminomethyl-4-mercaptopyrrolidine (3.3 Yield: Quantitative.

15 NMR 6 (CDC13) ppm: 1.63 to 1.90(m, 1H), 2.48 to 2.68(m, 1H), 2.86 to 3.43(m, 4H), 3.65 to 4.23(m, 2H), 5.22(s, 2H), 7.52(d, J=9Hz, 2H), 8.23 J=9Hz, 2H).

4 9* Step 6. Preparation of an N-sulfamoyl compound 20 To a solution of crude (2S,4S)-l-p-nitrobenzyloxycarbonyl-2-aminomethyl-4-mercaptopyrrolidine (3.3 g: 10.18 mmole) in dichloromethane (100 ml) at -78*C, triethylamine (2.84 ml: 2.2 eq.) and trimethylchlorosilane (3.12 ml: 2.2 eq.) are dropwise added.

After stirring for 20 minutes, triethylamine (4.25 ml: 3 eq.) and IM-sulfamoyl chloride in dichloromethane ml: 2.5 eq.) are dropwise added to the mixture.

After 20 minutes stirring, the reaction mixture is acidified with hydrochloric acid, warmed to room temperature, and extracted with dichloromethane. The extract is washed with water, and IN-hydrochloric acid ml) and methanol (30 ml) are added thereto. The solution is stirred at room temperature for 30 minutes.

47 The reaction mixture is washed with water, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate) to give (2S,4S)-l-p-nitrobenzyloxycarbonyl-2-sulfamoylaminomethyl -4-mercaptopyrrolidine (2.65 Yield: 66.7%.

NMR 6 (CDCl 3

-CD

3 0D) ppm: 1.42(t, J=7Hz, LH), 1.65 to 1.93(m, 1H), 2.48 to 2.70(m, 1H), 3.05 to 3.63(m, 4H), 3.93 to 4.16(m, 2H), 5.22(s, 2H), 7.53(d, J=8Hz, 2H), 8.23(d, J=8Hz, 2H).

IR (CHCl 3 cm- 1 3276br, 1692, 16C7.

PreparativeExample 4 of a pyrrolidine derivative

S

*S i 4r In.

.4 MsO.

N cx 1 Boc "cS f I 7 Boc HOE. a 0

H

.mou Boo tlsO~K~T Y Boo BoC 2 01.

3 Bo Boo Step 1. Preparation of a Boc compound To a suspension of trans-4-hydroxy-L-proline g) in a mixture of water (300 ml) and t-butanol (100 ml) are added aqueous sodium hydrogen carbonate (32.3 di-t-butyl dicarbonate (104 g) and dioxane 48 (200 ml). The mixture is stirred at room temperature overnight. The organic solvent is removed and the resulting aqueous solution is layered with methyl ethyl ketone and ethyl acetate, and acidified with conc.

hydrochloric acid (34.5 ml) under ice cooling. The organic layer is taken, washed with saturated brine, dried over sodium sulfate, and concentrated in vacuo.

The residue is recrystallized from ethyl acetate-toluene to give trans-l-t-butoxycarbonyl-4-hydroxy-Lproline (82.9 Colorless crystals. Yield: 94%.

mp. 126 to 128 0

C.

NMR 6 (CDC1 3 ppm: 1.43, 1.46(2 x s, 9H), 1.95 to goes 2.36(m, 2H), 3.36 to 3.6(m, 2H), 4.23 to 4.44(m, 2H).

IR (CHC13) cm- 1 3360, 1735, 1656.

15 Elemental analysis (C 10 H 7

NO

5 Calcd.: C, 51.94; H, 7.41; N, 6.06.

Found C, 51.65; H, 7.38; N, 5.99.

t4 Step 2. Preparation of a compound having mesyloxy and 20 methoxycarbonyl groups *m To a solution of trans-l-t-butoxycarbonyl-4hydroxy-L-proline (8.5 g) in tetrahydrofuran (110 ml) at -30 0 C, triethylamine (12.8 ml) and methanesulfonyl chloride (6.27 ml) are added. The mixture is stirred at the same temperature for 30 minutes. To the mixture triethylamine (5.13 ml) and methanol (30 ml) are added.

The mixture is stirred for 30 minutes. The reaction mixture is acidified with IN-hydrochloric acid (37 ml) and extracted with ethyl acetate. The extract is successively washed with water, aqueous sodium hydrog.n carbonate, water and saturated brine, dried over sodium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography and re- 49 crystallized from toluene-petroleum ether' to give (2S, 4R)-1-t-butoxycarbonyl-4-methanesulfonyloxypyrrolidine-2-carboxylic acid methyl ester (9.16 Colorless crystals. Yield: 77%. mp. 86 to 87'C.

NMR 8 (CDCl 3 ppm: 1.42, 1.47, 1.50(3 x s, 9H), 2.19 to 2,35(m, 1H), 2.48 to 2 .75(m, 3.06, 3.07, 3.26(3 x s, 3H), 3.59 to 4.12(m,, 5H), 4.35 to 4.60(m, 1H), 5.18 to 5.32(m, 1H-).

IR l( (CHCl 3 cm- 1 1748, 1698.

Step 3. Preparation of a methylol compound too.

To a solution of (2S,4R)-1-t-butoxycarbonyl- 4-methanesulfonyloxypyrrolidine-2-carboxylic acid methyl ester (8.11 g) in tetrahydrofuran (49 ml) stir- 15 ring under ice cooling, sodium borohydride (2.36 g) and methanol (20 ml) are added. The mixture is stirred at goes: room temperature for 25 minutes and at 60*C for #09 25 minutes. The mixture is cooled with ice and filtered. The filtrate is concentrated, diluted with 20 ethyl acetate, washed with water, dried over sodium sulfate, and concentrated in vacuo. The residue is recrystallized from petroleum ether-ether to give 99 9 (2S, 4R t-butoxycarbonyl-4-methanesulfonyloxypyrrolidine-2-methanol (5.96 Colorless crystals. Yield: 80%. mp. 95 to 96*C.

NMR 6 (CDCl 3 ppm: 1.48(s, 9H), 1.78 to 2.02(m, 1H), 2.3 to 2.48(m, 1H), 3 .05(s, 3H), 3.5 to 3.65(m, 2H), 3.65 to 4.0(m, 2H), 4.03 to 4.25(m, 5.2(s 1H-).

IR (CHCl 3 cm- 3460, 1680.

Step 4. Preparation of a tosyl compuund To a solution of (2S,.4R)-I-t-butoxycarbonyl- 4 -methanesulf onyloxypyrrol idine- 2-methanol (12.0 g) in 50 dichloromethane (180 ml) stirring under ice cooling, triethylamine (6.23 ml), p-toluenesulfonyl chloride (8.52 g) and N,N-dimethylaminopyridine (993 mg) are successively added. The mixture is heated to reflux for 3 hours, supplemented with triethylamine (0.57 ml) and p-toluenesulfonyl chloride (775 mg), and heated to reflux for 1 hour. The reaction mixture is acidified with dilute hydrochloric acid. The organic layer is taken, washed with water, dried over sodium sulfate and concentrated in vacuo. The residue is purified by silica gel column chromatography and recrystallized S' «from n-hexane to give (2S,4R)-l-t-butoxycarbonyl-4meth&nesulfonyloxypyrrolidine-2-methanol p-toluenesulfonate (16.8 Yield: 92%. mp. 65 to 66°C.

15 NMR 6 (CDC 3 ppm: 1.42(s, 9H), 2.15 to 2.55(m, 2H), 2.45(s, 3H), 3.03(s, 3H), 3.3 to 4.5(m, 5H), 5.1 to 5.25(m, 1H), 7.35(d, J=8.0Hz, 2H), 7.76(d, J=8.OHz, 2H).

IR (CHC13) cm- 1 1693.

Step 5. Preparation of a phthalimido compound To a solution of (2S,4R)-l-t-butoxycarbonyl- 4-methanesulfonyloxypyrrolidine-2-methanol p-toluenesulfonate (20.78 g) in dimethylformamide(200 ml), potassium phthalimide (9.61 g) is added. The mixture is stirred at 70°C for 3 hours. The reaction mixture is poured into a mixture of water and ethyl acetate.

The organic layer is taken, successively washed with dilute aqueous sodium hydroxide and water, dried over sodium sulfate, and concentrated in vacuo. The residue is purified by 5% wet silica gel column chromatography to give (2S,4R)-l-t-butoxycarbonyl-2-phthalimidomethyl- 4-methanesulfonyloxypyrrolidine (11.17 Yield: YI ~I II 51 Colorless foam.

NMR 6 (CDC1 3 ppm: 1.33, 1.42(2 x s, 9H), 2.0 to 2.55(m, 2H), 3.02(s, 3H), 3.4 to 4.6(m, 5H), 5.15 to 5.3(m, 1H), 7.6 to 7.95(m, 4H).

IR (CHC13) cm- 1 1775, 1716, 1693.

Step 6. Preparation of an acetylthio compound To a solution of (2S,4R)-l-t-butoxycarbonyl- 2-phthalimidometh 4-myl-4-methanesulfonyloxypyrrolidine (3 g) in dimethylformamide (30 ml), potassium thioacetate (1.65 g) is added. The mixture is stirred at for 3.5 hours. The reaction mixture is poured into a mixture of ethyl acetate and dilute hydrochloric acid.

The organic layer is taken, washed with water, dried 15 over sodium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography to give (2S,4S)-1-t-butoxycarbonyl-2-phthalimidomethyl-4-acetylthiopyrrolidine (2.12 Yield: 74%.

Orange colored syrup.

20 NMR 8 (CDC13) ppm: 1.30, 1,39(2 x s, 9H), 1.6 to S 2.0(m, 1H), 2.34(s, 3H), 2.4 to 2.67(m, 1H), 3.15 to 3.3(m, 1H), 3.65 to 4.55(m, 5H), 7.6 to 8.0(m, 4H).

IR V (CHC1 3 cm- 1 1774, 1715, 1688.

Step 7. Removal of a phthalyl and an acetyl groups To a solution of (2S,4S)-l-t-butoxycarbonyl- 2-phthalimidomethyl-4-acetylthiopyrrolidine (8.58 g) in a mixture of dichloromethane (26 ml) and methanol (129 ml), hydrazine hydrate (4.11 ml) is added. The mixture is he, ted to reflux for 2 hours and 45 minutes and filtered. The filtrate is concentrated in vacuo.

The residue is dissolved in dichloromethane, washed with water, dried over sodium sulfate, and concentrated eB~ 52 in vacuo to give crude (2S,4S)-l-t-butoxycarbon'yl-2aminomethyl-4-mercaptopyrrolidine (4.1 Yellow syrup.

Step 8 Preparation of a sulfamoyl compound To a solution of crude (2S,4S)-l-t-butoxycarbonyl-2-aminomethyl-4--mercaptopyrroliuine (4.1 g) in dichioromethane (250 ml) at -70'C und r a stream of nitrogen, triethylamine (8.87 ml) arid trimethylchlorosilarie (6.73 ml) are added. The mixture is stirred for 1 hour and 40 minutes 1 mixed with triethylamine (8.87 ml) and a solution of 1M-sulfamoyl chloride in dichioromethane (64 and stirred for 1 hour. The reaction mixture is acidified with dilute hydrochloric 15 acid. The organic layer is taken, diluted with lN-hydroc loric acid (21 ml) and methanol (50 ml), stirred for 35 minutes at room temperature, and poured Cinto water. The organic layer is taken, washed with watei dried over magnesium sulfate, and concentrated 20 in vacuo. The residue is purified by silica gel column a chromatography to give (2S, 4S )-l-t-butoxycarbonyl-2sulfamoylaminomethyl-4-mercaptopyr~rolidizhe (4.57 g).

Yield: 69%. ',lofless syrup.

NMR (CDCl 3 ppm: 1.46(s, 9H), 1.5 to 1.8(m, l1H), 1l1.71(d, .=6.6Hz, 1H), 2.5 to 2.67(m, 1HI), 3.0 to 3.46(m, 4H), 3.85 to 4.2(m, 2H), 4.6 (br s, 2H).

IR Q (CHC1 3 cm- 1 3420, 3340, 3270, 1679.

IUI~I rarm ~~rrra 53 Preparative Example 5 of a pyrrolidine derivative Ms0.. Ms.. Ms0..

11-COMe 1 2 3 Pmz Pmz

P

m z MsO&. Ac &Z N f HN NFt N- 4 i Pmz Pmz Pmz 2

NH

2 Pmz Step. 1 Preparation of a methylol compound Sa To a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-methanesulfonyloxypyrrolidine-2-carboxylic acid methyl ester (79.4 g: 205 mmole) in a mixture 0*'6 of tetrahydrqfuran (200 ml) and ethanol (300 ml), sodium borohydride (14 g) is added in several portions Sunder ice cooling. The mixture is stirred at room temperature for 4 hours. 'The reaction mixture is neutralized with conc. sulfuric acid, concentrated in vacuo to approximately a half volume, diluted with water, and extracted with ethyl acetate. The extract is successively washed with aqueous sodium hydrogen carbonate, water and brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate 1:2) to give (2S,4R)-l-p- 54 methoxybenzyloxycarbonyl-4 -methanesulfonyloxypyrrolidine-2-methanol (58.7 Yield: 81.7%.

NMR 6 (CDCl 3 ppm: 1.8 to 2.2(m, 1H), 2.3 to 1H), 3.01L(s, 3H), 3.57 J=4.4Hz, 111), 3.64(d, J=4.4Hz, 1H), 3.81(s, 3H1), 3.82 to 4.3(m,3H), 5.09(s, 2H), 5.21(br s, 1H), 6.89(d, J=8.8Hz, 2H), 7.31(d, J= 8.BHz, 2H) Step 2. Preparation of a tosyl compound To a solution of (2S,4R).-l-p-methoxybenzyloxycarbonyl- 4-methainesul fonyloxypyrrolidine-2-methanol (8.7 g: 24.2 mmole) in dichloromethane(80 ml) are added V"009triethylamine (4.05 ml), p-toluenesulfonyl chloride (5.08 g) and 4-dirnethylaminopyridine (148 mg). The 6 a0 0 15 mixture is stirred at room temperature overnight. The reaction mixture is washed with water arid brine, dried over magnesium sulfate, and concentrated in *.vacuo. The residue is purified by silica gel column chromatography (toluene ethyl acetate 1 1) to v.

00 0. 20 give (2S,4R)-1-p-methoxybenzyloxycarbonyl-4-methanesulfonyloxypyrrolidine-2-methanol p-toluenesulfonate (11.75 Yield: NMR (CDC1 3 ppm: 2.2 to 2.44(s, 3H1), 0 0 6 0 a2.98(s, 3H), 3.4 to 3.6(m, 2H), 3.82(s, 311), 3.8 to 5.03, 4.95(ABq, 3=12Hz, 211), 5.2 (br s, 1H1), 6.89(d, 3=8.6Hz, 2H1), 7.18 to 7.4(m, 4H), 7.6 to 7.8(m, 2H).

IR Q (CR01 3 cm- 1 1698, 1612.

Step 3. Preparation of a phthalimido compound To a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-methanesul fonyloxypyrrolidine-2-methanol p-toluenesulfonate (6.35 g: 12.27 mmole) in dimethyl- 55 formamide (60 ml), potassium phthalimide (2.7 g) is added. The mixture is stirred at 7000 for 4 hours.

The reaction mixture is poured into ice water and extracted with ethyl acetate. The extract is successively washed with water and brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluene: ethyl acetate 2:1) to give (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-methanesulfonyloxy-2-phthalimidomethylpyrrolidine (4.65 Yield: 77.5%.

NMR 5 (CDCl 3 ppm: 2 to 2.3(m, 1H), 2.4 to 2.6(m, lH), 2.95, 2.97(2 x s,3H), 3.43 to 4.2(m, 5H), 3.80(s, 3H), [5.01(s)+i5.07, 4.96(ABq, 12.2Hz), 2H], 5.13 to 5.3(mn, 1H).

IR (CHCl 3 cm- 1 1774, 1716, 1613.

~:'.Step 4. Preparation of an acetylthio compound To a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl-4 -methanesulfonyloxy- 2-phthalimidopyrrolidine (4.0 g: 8.19 mmole) in dimethylformamide (40 ml), potassium thioacetate (2.1 g) is added. The mixture is stirred at 600C for 3 hours. The reaction mixture is diluted with ethyl acetate, successively washed with water and brine, dried over magnesium sufate, and concentrated in vacuo. The residue is p -:ified by silica gel column chromatography (toluene :ethyl acetate to give (2S,4S)-l-p-methoxvbe.zvloxycarbonyl-4-acetylthio-2-phthalimidomethyip,,rrolidine (3.2 Yield: 78%, NMR iS (CDCl 3 ppm: 1.7 to 1.9(m, 1H), 2.4 to 2.7(m, 1H), 3.21, 3.26(2 x d, J=7Hz, 3.8(s, 3H), 3.7 to 4.2 to 4.5(m, 1H), [4.95(s) +5.04, 4.83(A~q, J=l2Hz), 2H], 6.83(d, J=7.6Hz, 2H), 7.18 to 7.3(m, 2H), t 56 7.6 to 7.9(m, 4H).

IR (CHC1 3 cm- 1 1773, 1714.

Step 5. Removal of an acetyl and a phthalyl groups and introduction of a sulfamoyl group To a solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl-4-acetylthio-2-phthalimidomethylpyrrolidine (4.3 g: 9.18 mmole) in a mixture of dichloromethane '13 ml) and methanol (65 ml), hyruzine hydrate (1.78 ml) is added. The mixture is heated to reflux for 4 hours. The reaction mixture is concentrated in vacuo. The residue is dissolved in dichloromethane and S filtered under a stream of nitrogen to remove solid.

The filtrate is washed with water, dried over magnesium 15 sulfate, and concentrated in vacuo. The resulting residue containing (2S,4S)-l-p-methoxybenzyloxycarbonyl-4-mercapto-2-aminomethylpyrrolidine is diluted with dichloromethane (100 ml), added triethylamine (2.63 g) and trimethylchlorosilane (2.4 ml) at -78 C, and 20 stirred for 20 minutes. To the reaction mixture, triethylamine (2.63 ml) and a solution (16.5 ml) of 31-sulfamoyl chloride in dichloromethane are added.

The mixture is stirred for 20 minutes. The reaction mixture is washed with dilute hydrochloric acid, mixed with 1N-iiydrochloric acid (9 ml) and methanol (20 ml), and stirred at room temperature for 30 minutes. The reaction mixture is successively washed with water and brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography (toluen ethyl acetate 1:2) to give (2S,4S)-l-p-methoxybenzyloxycarbonyl-4-mercapto-2-sulfamoylaminomethylpyrrolidine (2.71 Yield:78.6%.

57 NMR (COJ- 3 ppm: 1.6 to 2.O(m, 2H), 2.4 to 2.7(m, 1H), 3.1 to 3.8(m,4H), 3.81(s, 3H), 3.9 to 4.2(m, 21), 4.6 to 5.O(m, 2H), 5.04(s, 2H), 5.97(br s, 11), 6.89(d, 3=8.6Hz, 2H), 7.30(d, J=8.6Hz, 2H).

IR (CHCl 3 cm- 3668, 3424, 1683.

Preparative Example 6 of a pyrrolidine derivative .9 9 a 9 9* 11 N Prm Ho Pm2 110.. 3-1j 4-1 PmZ 11'.'7KN152N12 -3 1 Pmz H0 3-2 110( .II 7N ,-Rrc -4-2 X~ KP Nrcy 3-1 4-1i 3-23 4 Prc NPrc NFt 4-31

INKNH

2 4 Pm 9 a.

a *a .9 6 7.i%,,NHS 2

NH

2 Pmz Pnz~ Step 1. Preparation of a methylol compound To a solution of trans-1-p-methoxybenzyloxycarbonyl-4-hydroxyproline (105.5 g: 357.5 mmole) in tetrahydrofuran (1 liter), triethylamine (54.8 ml) is added. After adding ethyl chloroformate (35.9 ml) dropwise at -30'C, th mixture is stirred for 58 minutes. To the reaction mixture, a solution of sodium borohybride (33.25 g) in water (120 ml) is added dropwise at a temperature in the range of -15 to neutralized with conc. hydrochloric acid, and concentrated in vacuo. To the residue, ethyl acetate is added, washed with brine, dried over magnesium sulfate, and concentrated in vacuo to give (2S,4R)l-p-methoxybenzyloxycarbonyl-4-hydroxy-pyrrolidine-2methanol.

NMR 6 (CDCl 3 PPM: 1.6 to 1.8(m, 1H), 1.95 to 2.2,m, 1H), 3.4 to 3.8 (in, 4H), 3.8(s, 3H), 4.0 to 4.3(m, 1H), 4.37(br s, 1H), 5.07(s, 2H), 6.88(d, J=8.8Hz, 2H), 7.30(d, J=8.8H-z, 2H).

to. 15 Step 2. Preparation of a tosyl compound 0: To a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl 4-hydroxypyrro Iid ine-2-methanol 64 g: 227.6 minole) in pyridine (350 ml), p-toluenesulfonyl **chloride (48 g) is added. The mixture is stirred at a as 20 room temperature for 4 hours. The reaction mixture is ago* poured into ice water and extracted with ethyl acetate.

The extract is successively washed with water, dilute hydrochloric acid and aqueous sodium hydrogen carbonate, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel column chromatography to give (2S, 4R)-l-p-methoxybenzyloxycarbonyl-4 -hydroxypyrrolidine- 2-methanol p-toluenesulfonate (60 g).

NMR 6 (CDCl 3 ppm: 2.0 to 2.4(m, 2H), 2.44(s, 3H), 3.3 to 3.7(m, 2H), 3.82(s, 3H), 3.9 to 4.6(m, 4H), 4,8 to 5.3(mn, 2H), 6.88(d, J=8.6Hz, 2H-),7.2 to 7.4(m, 4H), 7.6 t 7.8(m, 2H-).

IR '2 (CHC1 3 cm- 1 3446, 1693.

59 Step 3. Preparation of an amino precursor (-NProc azido) 1) To a solution of (2S,4R)-l-p-methoxybenzyloxycarbonyl-4--hydroxypyrrolidine-2-methanol p-toluene- (8.7 g: 20 mmole) in dimethylformamide ml), a solution of sodium azide (1.56 g) in water (6 ml) is added. The mixture is stirred at 80 0 C overnight. The reaction mixture is poured into ice water and extracted with ethyl acetate. The extract is :0000 10 successively washed with water and brine, dried over off.*magnesium sulfate, and concentrated in vacuo to give crude (2S, 4R)-l--p-methoxybenzyloxycarbonyl-4--hydroxy-2azidomethylpyrrolidine (5.6 Yield: 92%.

*got NMR 5 (ODC1 3 ppm: 1.95 to 2.1(m, 2H), 3.2 to 3.8(m, 3H), 3.81(s, 3H),3.83 to 4.6(m, 3H), 5.07(s, 2H4), 6.89(d, J=8.8Hz, 2H), 7.31(d, J=8.8Hz, 2H).

IR \)(CHCl 3 cm- 1 3420, 2100, 1689.

2 To a solution of the thus obtained 20 (2S, 4R)-l-p-methoxybenzyloxycarbonyl-4-hydroxy-2-azidomethylpyrrolidine (5.57 g: 18.18 mmole) in methanol ml) are added, 5% palladium on charcoal (560 mg) 6 0 0and ammonium formate (2.3 The mixture is stirred at 45'C for 2 hours. The reaction mixture is diluted with dichioromethane (50 ml), filtered to remove the catalyst, and concentrated in vacuo. The residue is crystallized from a mixture of dichioromethane and ether and washed with ether to give (2S,4R)-1--pmethoxcybenzyloxycarbonyl-4 -hydroxy-, 2-aminomethylpyrrolidine formate (4.28 Yield: 72%.

NMF. 6 (CDCl 3

-CD

3 OD) ppm: 1.6 to 1,82(m, 1H), 2.1 to 2.3(m, 1H), 2.7 to 3.7(m 4H), 3.81(s, 4.1 to 4.4(m, 2H), 5.04(s, 2H), 6.88(d, 3=8.8 Hz, 2H), 7.28(d, 60 3=8.8Hz, 2H), 8.47(s, J.H).

IR (CHCl 3 cm- 1 3678, 3412, 1678, 1602.

Step 4. Introduction of a protected amino group, removal of the protection, and introduction of a sulfamoyl group (-NProc phthalimido) 1) To a solution of (2S,4R)-1-p-methoxybenzyloxycarbonyl 4- hydroxypyrrolidine- 2-methanol p-toluenesulfonate (24 g: 55.1 mmole) in dimethylformamide V04 10 (200 ml), potassium phthalimide (15.3 g) is added. The mixture is stirred at 80*C for 4 hours. The reaction mixture is diluted with ethyl acetate, successively Sos washed with water and brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is 15 purified by sil~ica gel column chromatography (toluene ethyl acetate 1:2) to give (2S,4R)--p-methoxybenzyloxycarbonyl..4-hydroxy-2-phthalimidomethylpyrroli- 9dine (18.1 Yield: NMR 5 (CDCl 3 ppm: 1.9 to 2.2(m, 2H), 3.4 to 4.05(m, 20 5H), 3.80(s, 3H), 4.3 to 4.6(m, 2H), 4.8 to 5.1(m, 2H), off. 6.83(d, J=8.2Hz, 2H), 7.25(d, 3=8.2Hz, 2H), 7.6 to 7.9(m, 4H-).

IR Y (CHCl 3 cm: 3458, 1773, 1712.

93) cm- 2) To a solution of (2S,4R)-l-p-methoxybenzyloxycarboriyl- 4-hydroxy-2 -phthalimidomethylpyrrolidine (5.13 g: 12.5 mmole) in a mixture of dichloromethane ml) and methanol (50 ml), hydrazine hydrate ml) is added. The mixture is heated -to reflux for 2 hours and concentrated in vacuo. The residue is diluted with dichloromethane, filtered to remove solid, washed with brine, dried over magnesium sulfate, and concentrated in vacuo to give a residue containing 61 (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-hydroxy-2-aminomethylpyrrolidine as a main component.

3) To a solution of the above-mentioned residue in dichloromethane (70 ml) at -70*C, triethylamine (4.6 ml) and trimethylchlorosilane (3.7 ml) are ,added. The mixture is stirred for 20 minutes. To the reaction mixture, triethylamine (5.5 ml) and a solution of 1M-sulfamoyl chloride in dichloromethane (34 ml) 10 are added. The mixture is stirred for 15 minutes.

The reaction mixture is washed with dilute hydrochloric acid, mixed with methanol (50 ml), and then 4Nhydrochloric acid (3.3 ml) is added under ice o cooling. After stirring the mixture, aqueous sodium hydrogen carbonate is added. The organic layer is taken, washed with water and brine, dried over magnesium sulfate, and concentrated in vacuo to give crude (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-hydroxy-2sulfamoylaminomethylpyrrolidine (3.96 g).

S 20 NMR 6 (CDC13) ppm: 1.8 to 2.25(m, 2H), 3 to 7H), 3.79(s, 3H), 5.03(s, 2H), 5.2 to 5.8(m, 2H), 6.08(br s, 1H), 6.87(d, J=8.8Hz, 2H), 7.29(d, J=8.8Hz, 2H).

IR Y (CHC1 3 cm- 1 3456, 1689.

Step 5 Preparation of a mesyl compound To a solution of crude (2S,4R)-1-p-methoxybenzyloxycarbonyl-4-hydroxy-2-sulfamoylaminomethylpyrrolidine (1.8 g: 5 mmole) obtained in Step 4 in dichloromethane (20 ml) at -70 0 C, triethylamine (0.77 ml) and methanesulfonyl chloride (0.39 ml) are added. The mixture is stirred for 45 minutes. The reaction mixture is neutralized with dilute hydrochloric acid,

I

62 successively washed with water and brine, and concentrated in vacuo to give crude (2S,4R)-1-p-methoxybenzyloxycarbonyl -4 -methanesulfonyloxy- 2- sul famoylaminomethylpyrrolidine (2.26 g).

NMR 8 (CDCl 3 ppm: 2 to 2.5(m, 2H), 2.99(s, 3H), to 4.3(m, 5H), 3.79(s, 3H), 4.8 to 5.3(m, 3H), 5.05(s, 2H), 5.7 to 5.85(m, 1H), 6.88(d,J=8.8Hz, 2H), 7.29(d, 3=8.8Hz, 2H).

IR (CHC2L 3 cmf 1 3606, 3416, 1690.

Step 6. Preparation of an acetylthio compound To a solution of crude (2S,4R)-l-p-methoxybenzyloxycarbonyl-4-methanesul fonyloxy- 2-sulfamoylaminomethylpyrrolidine (2.26 g) obtained in Step 5 in 15 dimethylformamide (12 ml), potassium thioacetate (1.7 g) is added. The mixture is stirred at 60'C for hours. The reaction mixture is diluted with ethyl acetate, success~ively wash _d wi th water and brine, dried over magnesium sulfate, and concentrated in vacuo. TbR residue is purified by silica gel column chromatography (toluene ethyl acetate =1 2) to give (2S,45 )-l-p-methoxybenzyloxycarbonyl-4-acetylthio- 2-sulfamoylaminomethylpyrrolidine (971 mg).

NMR 6 (CDC1 3 ppm: 1.8(br s, 1H), 2.33(s, 3H), 2.4 to 2.7(m, 1H), 3.1 to 3.81(s, 3H), 3.9 to 4.2(m, 2H), 5.05(s, 2H), 6.89(d, 3=8.8Hz, 2H), 7.30(d, 3=8.8Hz, 2H).

IR (CHCl 3 cm- 3414, 3276, 1688.

Step 7. Removal of an acetyl group To a solution of (2S,4S)-l-p-methoxybenzyloxycarbonyl-4-acetylthio-2-sulfamoylaminomethylpyrrolidine (982 mg: 2.35 mmole) in a mixture of dichlorometh- 63 ane (2 and methanol (10 ml), 1N-sodium hydroxide (2.8 mnl) is added under ice cooling. The mixture is stirred for 15 minutes. The reaction mixture is diluted with water and extracted with ethyl acetate. The extract is successively washed with water and brine, dried over magnesium sulfate, and concentrated in vacua. The residue is purified by silica gel column chromatography (toluene ethyl acetate 2) to give (2S,4S)-l-p-methoxybenzyloxyc.3rbonyl-4-mercapto-2sulfamoylaminomethylpyrrolidine (783 mng). Yield: 89%.

~NMR .5 (CDCl 3 pm 1.6 to 2.0(m, 2.4 to 2.7(m, 3.1 to 3.8(m, 4H), 3.81(s, 3H), 3.9 to 4.2(m, 2H), so of 4.6 to 5.0(m, 2H), 5.04(s, 2H), 5.97(br s, 1 H), 6.89(d, 3=8.6Hz, 2H), 7.30(d, J=8.6Hz, 2H).

&go 15 IR (CR01 3 cm- 1 3668, 3424, 1683.

0: 9.

Preparative Example 7-A of a pyrrolidine derivative HO. H..,H 11 HO.,2.

ODI2OH HO..,01l *H H HC1 Bo lhI O 6 Ac-S-. Boc DoC N "'SO 2

E'

2 N-S2H 64 Step A-1. Preparation of an ester compound To a suspension of cis-4-hydroxy-D-proline (16.46 g: 125.5 mmole) in methanol (66 ml), thionylchloride (9.16 ml: 125.5 mmole) is added in a nitrogen atmosphere under ice cooling, and the mixture is stirred at room temperature for 30 minutes. The mixture is further stirred to react at 40*C for 4 hours to give (2R,4R)-4-hydroxy-2-methoxycarbonylpyrrolidine hydrochloride as crude crystals (25.74 Yield: 113 Colorless crystals.

SNMR 6 (D 2 0) ppm: 2.3 to 2.6(m, 2H), 3.33(s, 1H), 3.4 to 3.5(m, 2H), 3.84(s, 3H), 4.6 to 4.7(m, 2H).

S" IR (KBr) cm- 1 3320, 2980, 1728.

15 Step A-2. Preparation of a Boc compound To a suspension of (2R,4R)-4-hydroxy-2methoxycarbonylpyrrolidine hydrochloride (25.64 g: 125 mmole) in dichloromethane (125ml), triethylamine (19.11ml: 137.5 mmole) is added dropwise in a nitrogen 20 atmosphere under ice cooling. The mixture is stirred for 5 minutes at room temperature. Then, a solution of di-t-butyl dicarbonate (34.11 g: 156.3 mmole) in dichloromethane (125 ml) is added dropwise, and the mixture is stirred for 40 minutes at room temperature to give (2R,4R)-l-t-butoxycarbonyl-4-hydroxy-2-methoxycarbonylpyrrolidine (26.85 Yield: 88%. Colorless crystals.

NMR 6 (CDC13) ppm: 1.46(d, J=8.4Hz, 9H), 2.0 to 2.2(m, 1H), 2.2 to 2.5(m, 1H), 3.4 to 3.8(m, 2H), 3.79(d, J=3.0Hz, 3H), 4.2 to 4.5(m, 2H).

IR Yl (KBr) cm- 1 3460, 1730, 1680.

65 Step A-3. Preparation of a mesyl compound To a solution of (2R,4R)-1-t-butoxycarbonyl- 4-hydroxy-2-methoxycarbonylpyrrolidine (9.81 g: mmole) in dichlorow~ethane (49 ml) in a nitrogen atmosphere under ice cooling, triethylamine (6.67 ml: 48 mmole) and methanesulfonyl chloride (3.70 ml: 48 mmole) are added. The mixture is stirred for minutes to give (2R,4R)-l-t-butoxycarbonyl-4methanesulfonyloxy-2-methoxycarbonylpyrrolidine as a crude oil (13.05 Yield: 101%.

NMR 5 (CDCl 3 PPM: 1.46(d, J=9.6Hz, 9H), 2.5(m, 2H), 3.02(s, 3H), 3.76(s, 3H), 3.8(m, 2H), 4.3 to 4.6(m, 1H), 5.2 to 5.3(m, 1-H).

Step A-4. Preparation of a methylol compound To a solution of (2R,4R)-l-t-butoxycarbonyl- 4-methanesulfonyloxy-2-methoxycarbonylpyrrolidirne 0 (11.21 g: 34.4 mmole) in a- mixture of tetrahydrofuran (34 ml) and ethanol (51 ml), sodium borohydride *0 20 (5.21 g: 137.7 mmole) is added in a nitrogen atn sphere under ice cooling. The mixture is stirred for minutes at room temperature to give (2R,4R)-l-t- ~butoxycarbonyl -4-methanesul fonyloxypyrrolidine- 2-metha nol (8.47 Yield: 83%. Colorless crystals.

NMR .6 (CDCl 3 ppm: 1.48(s, 9H), 1.9 to 2.2(m, 1H), 2.3 to 2.5Cm, 1H), 3.06 3H), 3.65(dd, J=1l.2Hz, 3J=4.0Hz, IH), 3.5 to 3-9(m, 2H), 3.84(dd, J-'ll.2Hz, J=7.6Hz, lH), 4.1(m, 1H), 5.2(m, 1H).

IR (KBr) cm- 1 3490, 1688.

Step A-5. Preparation of an acetylthio compound (2R, 4R )-l-t-Butoxycarbonyl-4-metha-nesulfonyloxypyrrolidine-2-methan)! a substrate) and 66 potassium thioacetate (KSAc) are dissolved in dimethylformamide (DMF), and the mixture is stirred. The conditions for this reaction are shown in Table 1, Step The reaction mixture is diluted with ethyl acetate, and ice water is added. The organic layer is taken, successively washed with aqueous sodium hydroxide, hydrochloric acid, water and saturat, 1 brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel chromatography to give (2R,4S)-4-acetylthio-l-t-butoxycarbonylpyrrolidine-2-methanol.

NMR 6 (CDC1 3 ppm: 1.47(s, 9H), 2.05(t, 2H), 2.34(s, 3H), 3..0 to 3.3(m, 1H), 3.40(dd, J=11.6Hz, J=5.2Hz, seed 1H), 3.5 to 3.9(m, 3H), 3.9 to 4.2(m, 2H).

Step A-6. Introduction of a sulfamide group a) Production of N-t-butoxycarbonylsulfamide so0: A solution of t-butanol (4.72 ml: 50 mmole) in ethyl acetate (100 ml) is cooled to -40 0 C, chlorosulfonyl 9* 20 isocyanate (4.35 ml: 50 mmole) is dropwise added thereto, and the mixture is stirred at -18°C for 20 minutes.

The reaction mixture is cooled to -72 C, gaseous ammonia (2 mole) is bubbled with stirring, and the mixture is stirred for 50 minutes while warming up to The reaction mixture is acidified with acid (30 ml) and the formed precipitate is filtered off. The organic layer is taken, successively washed with water and brine, dried over magnesium sulfate, and concentrated under reduced pressure. The crystalline .residue is washed with hexane-ethyl acetate ml) and recrystallized from ethyl acetate-hexane to give N-t-butoxycarbonylsulf.mide (8.81 Yield: 89%.

Colorless crystals. mp.130 to 131'C.

67 NMR 8 (CD 3

SOCD

3 PPM 1.43(s, 7.27(s, 2H).

IR (NUjol) cm- 1 3360 3270, 1718, 1548.

Elemental Analysis (C 5 Hl 2

N

2 0 4

S)

Calcd.: C, 30.60; H, 6.17; N, 14.28; S, 16.34.

Found 30.39; H, 6.11; N, 14.30; S, 16.30.

b) Preparation of a sulfamide compound To a solution of (2R,4S)-4-acetylthio-1-t--butoxycarbonylpyrrolidine-2-methanol a substrate) in tetrahydrofuran (THF), triphenyiphosphine (PPh 3 N-t-butoxycarbonylsulfamide (BSMD), and azodicarboxylic acid diethyl ester (DEAD) are successively added under ice cooling. The conditions for this reaction are shown in Table 2, Step The reaction mixture is 15 diluted with toluene, concentrated, diluted with toluene, and the formed crystals are filtered off. The filtrate is concentrated. The residue is purified by silica gel column chromatography to give (2R,4S)-4acetylthio-l-t-butoxycarbonyl-2- (N-t-butoxycarbonyl-N- 20 sulfamoylamino)methylpyrrolidine.

NMR 5 (CDCl 3 pm 1.41(s, 9H), 1.55(s, 911), 1.9 to 2.0(m, 211), 2.35(s, 3H), 3.32(dd, 3=11.4Hz, OleJ3= 8. 2Hz, 1H) 3. 6 to 3. 9 3 H) 3. 9 to 4. 1(m, 1H) 6.15(s, 2H1).

.IR (KBr) cnf 1 3420, 3320, 1706, 1686, 1666.

Step A-7. Removal of an acetyl group To a solution of (2R,4S)-4-acetylthio-l-tbutoxycarbonyl-2-(N-t-butoxycarbonyl-N-sulfamoylamino) methylpyrrolidine a substrate) in dichloromethane, 4.92 M sodium methoxide (NaDMe) in mnethainol is added. The mixture is stirred. The conditions for this reaction are shown in Table 3, Step A-7. The 68 reaction mixture is diluted woith water. The water layer is taken, toluene is added thereto, and acidified with conc. ,ydrochJloric acid under ice cooling. The organic layer is taken, successively washed with water and saturated brine, dried over magnesium sulfate, and concentrated in vacuo to give (2R,4S)-l-t-butoxycarbonyl-2-(N-t-butoxycarbonyl-N-su2.famoylamino)methyl-4mercaptopyrrolidine. mp. 90.0 to 91.5*C.

NMR (ODd1 3 ppm: 1.43(s, 9H), 1.52(s, 9H), 1.72(d, J=7.0Hz, 1H), 1.9 to 2.0(m, 2H), 3.2 to 3.8(m, 0O@@ 4.5(m, 1H), 6.11(s, 2H).

IR 0) (KBr) cm- 1 3220, 1698, 1683.

:0:Elemental Analysis (C 15

H

29 0NS) Calcd. C:43.78, H:7.10, N:10.21, S:15.58.

Found. C:43.55, H:7.11, N:10.37, S:15.75.

Preparative Example 7-B of a pyrrolidine derivative

HO,...DD-

CMD CO Ot 4O, MS_ 2 3, .bc Bc Boc 6 AcS., Hoc BSI., BHoc CI' ."-SO'LNH 2 N "-SO 2 11 2 Hoc o 69 Step B-1. Substitution for a formylo yo~u To a solution of (2R,4R)-1-t-butoxycarbonyl- 4-hydroxy-2-methoxycarbofllpyrrolidine (2.45 g: mmole) in tetrahydrofuran (10 ml), formic acid (453 pi: 12 mmole), triphenyiphosphine (3.15 g! 12 mmole), and diet~hyl azodicarboxylate (1.89 ml: 12 mmole) are successively added in a nitrogen atmosphere under ice cooling. The mixture is stirred for 30 minutes at the same temperature to give (2R, 4S )-l-t-butoxycarbonyl-4-formyloxy-2methoxycarbonylpyrrolidine (2.17 Yield: 79%.

Colorless oil.

NMR 6 (CDCl 3 ppm: 1.44(d, J=7.8Hz, 9H), 2.1 -to 2.6(m, 2H), 3.5 to 3.9(m, 5H), 4.4(m, 1H), 5.4(m, 1H), ag* 15 8.0(s, 1H).

Step B-2. Removal of a formyl group To a solution of (2R,4S)-1-t-butoxycarbonyl- 4-formyloxy-2-methoxycarbonylpyrrolidine (2.08 g: a 20 7.6 mmole) in methanol (21.0 ml), aqueous lN-sodium hydroxide (7.6 ml) is added under ice cooling. The mixture is stirred at the same temperature for minutes to give (2R. 4S)-l-t-butoxycarbonyl-4hydroxy-2-rfethoxycarbonylpyrrolidine (1.86 g).

Yield: 100%. Colorless oil.

NMR 6 (ODC1 3 PPM: 1.44(d, J=9.2H-z, 9H), 1.9 to 2.4(m, 2H), 3.4 to 3.7(m, 2H), 3.74 3H), 4.3 to 4.6(m, 2H).

Step B-3. Preparation of a mesyl compound To a solution of (2R,4S)-1-t-butoxycarbonyl- 4-hydroxy-2-niethoxycarbonylpyrrolidile (3.17 g: 12.9 mmole) in dichloromethane, (16 ml) in a nitrogen 70 atmosphere under ice cooling, triethylamine (2.15 ml: 15.5 mmole) and methanesulfonyl chloride (1.19 ml: 15.5 mmole) are added. The mixture is stirred to react for 30 minutes to give (2R,4S)-l-t-butoxycarbonyl-4methanesulfonyloxy-2-methoxycarbonylpyrrolidine as oil (4.13 Yield: 99%.

NMR 6 (%CDC1 3 ppm: 1.46(d, J=8.4Hz, 9H), 2.3(m, 1H), to 2.8(m, 1H), 3.08(s, 3H), 3.8 to 4.0(m, 5H), 4.3 to 4.6(m, 1H), 5.3(m, 1H).

cC, Step B-4. Preparation of a methylol compound To a solution of (2R,4S)-l-t-butoxycarbonyl- 4-methanes u f onyloxy -2-met ho xycarbonyIp yrro lidime (3.96 g: 12.2 mmole) in a mixture of tetrahydrofuran 15 (12 ml) and ethanol (18 ml), sodium borohydride (1.85 g: 48.8 mmole) is added in a nitrogen atmosphere under ice cooling. The mixture is stirred for C 45 minutes at room temperature to give (2R,4S)-l-tbutoxycarbonyl-4 -methanesulfonyloxypyrrolidine-2-metha- 20 nol (2.97 Yield: 83%. Colorless crystals. mp. to 96'C.

NMR 6 (CDCl 3 ppm: 1.49(s, 9H), 1.7 to 2.1(m, 1H), 2.3 to 2.5(m, 1H), 3.06(s, 3H), 3.4 to 3.7(m, 2H), 3.7 to 4.0(m, 2H), 4.0 to 4.3(m, 5.2(m, 1H).

IR 2 (KBr) cm- 1 3400, 3420, 1648.

Step B-S. Substitution for an acetylthio group (2R, 4S)-1-t-butoxycarbonyl-4-methanesulfonyloxypyrrolidine-2-methanol a substrate) is allowe~d to re.act in the same manner as in Step A-S in Preparative Example 7-A under a condition for Step B-S shown in Table 1 to give (2R,4R)-4-acetylthio-l-tbutoxycarbonylpyrrolidine-2-methanol.

-71 NMR 8 (C~DC 3 PPM: 1.47(s, 91-1), 2.34(s, 314), 2.4 to 3.2 (in, 2H), 3.58 to 4.1(m, 611).

iiR 0 (C1-Cl 3 cnf 1 3380, 1690.

Step B-6. introduction of a sulfamide group N-t-butoxycarbonylsulfamide is prepared in the same manner as in the paragraph of Step A-6 in Preparative Example 7-A. (2R,4R)-4-acetylthio-l-tbutoxycarbonylpyrrolidine-2-methanol a substrate) is allowed to react with the obtained N-tbutoxycarbonylsulfamide in the similar manner as in paragraph of Step A-6 in Preparative Example 7-A under a condition for Step B-6 shown in Table 2 to **give (2R,4f)-4-acetylthio-l-t-butoxycarbonyl-2-(N-tbutoxycarbonyl-N--sulfamcylamino )methylpyrrolidine.

NMR 8 (ODC1 3 ppm 1.43(s, 9H), 1.53(s, 9H), 2.34(s, 3H), 2.5(m, 1H), 3.15(dd, J=12.2Hz, J=6.2Hz, 1H), *se 3.58(dd, J=14.8Hz, J=3.2Hz, 1H), 3.8 to 4.1(m, 2H), 4.16(dd, J=12.2Hz, J=7.8Hz, 1H), 4.4 to 4.7 (in, 1H), 20 6.11(s, 2H).

IR (CHCl 3 cm- 1 3360, 3200, 1710, 1688.

Step B-7. Removal of an acetyl group 6 (2R, 4R)-4-acetylthio-l-t-butoxycarbonyl-2-(Nt-butoxycarbonyl-N-sulfamoylamino) methylpyrrolidine a substrate) is deacetylated in the similar manner as in Step A-7 of Preparative Example 7-A under a condition for Step B-7 shown in Table 3 to give (2R,4R)-l-t-butoxycarboniyl-2-(N-t-butoxycarbonyl-Nsulfamoylamino)methyl-4-mercaptopyrrolidine. mp. 92 to 93'C.

INMR 6 (CDC1 3 ppm :2to5(,11H), 1.42 9H), 1.54(s, 9H), 1.82(d, J=6.2H-z, 1H), 2.5 to 2.7(m, 1H1), 72 4.09, 3.05 (ABX, 3=12.0Hiz, J=7.414z, J=8.2Hz, 2H), 4.06, 3.62(ABX, Jt=15.0HZ, J=10.8 Hz, 3=3.2Hz, 2H), 4.2 to 4.6(m, 1H), 6.08(s, 2H1).

IR 0, (CHCl 3 cm- 1 3380, 3220, 1718, 1680.

Preparative Example 7-C of a pyrrolidine derivative

HO..,

NH

Boo H HG1 e 4 H'I e Boo AO

O~

b~ 1 c00~ 3 Boc see* Doe a 6 hs7 Boo Boo AcS.,,8 113S.., Boc Boc es so* *00.

Step C-1.Preparation of an ester compound To a suspension of trans-4-hydroxy-L-prol-Ine (200*g: 1.525 mole) in methanol (800 nil), acetylchlo- 10 ride (163 mil: 2.288 mole) is added dropwise under ice cooling in a nitrogen atmosphre. The mixture is warmed to room temperature, mixed with thionyl chloride (55.7 mil: 0.763 mole), and stirred for 4 hours at to give (2S, 4R)--yrx--mtoyabnypirldn hydrochloride (244.27 Yield: 88%. Colorless crystals.

I

73 NMR 6 (D 2 0) ppm: 1.8 to 2-0(m, 1H), 2.0 to 2.2(m, 1H), 2.9 to 3.1(m, lIH), 3.17(dd, 3=12.6Hz, 7=3.6Hz, 1H), 3.49(s, 3H), 4.2 to 4.4(m, 2H1).

IR (KBr) cm-l: 3380, 3330, 2695, 2960, 1742.

Step C-2. Preparation of a Boocomp ound To a suspension of (2S,4R)-4-hydroxy-2methoxycarbonylpyrrolidine hydrochloride (12.71 g: mmole) in dichioromethane (70 ml), triethylamine (10.7 ml: 77 mmole) is added dropwise under ice cooling in a nitrogen atmosphere. The mixture is stirred for min. at room temperature. A solution of di-t-butyl dicarbonate (19.10 g: 87.5 mmole) in dichioromethane (72 ml) is added dropwise thereto, and the mixture is stirred for 45 minutes at room temperature to give (2S, 4R) -l-t-butoxycarbonyl-4-hydroxy--2-methoxycarbonylpyrrolidine (14.06 Yield: 82%. Colorless oil.

NMR 6 (Dd 3 pp:1.44(d, J=9.6Hz, 9H1), 1.9 to 2.4(m, 3H1), 3.4 to 3.7(m, 2H), 3.74 311), 4.3 to 4.6(m, 2H).

Step C-3. Substitution with a formyloxy groiup To a solutio'n of (2S,4R)-l-t-butoxycarbonyl- 4-hydroxy--2-methoxycarbonylpyrrolidine (7.36 g: 25 30 mmole) in tetrahydrofuran (30m1), formic acid (1.36 ml: 36 rnmole), triphenyiphosphine (9.44 g: 36 mmole) and diethyl azodicarboxylate (5.67 ml: 36 mmole) are successively added in a nitrogen atmos- *phere under ice cooling. The mixture is stirred to react for 40 minutes at the same temperature to give (2S,4S )-l-t-butoxycarbonyl-4-formyloxy-2-methoxycarbonylpyrrolidine (5.38 Yield: 66%. Colorless crystals.

74 NMR 5 (ODC1 3 PPM: 1.45(d, 3=8.6Hz, 9H), 2.2 to 2.4(m, 1H), 2.4 to 2.7(m, 1H), 3.5 to 3.9(mn, 2H), 3.75(s, 3H), 4.3 to 4.6(m, 1H), 5.3 to 5.5(m, 1H), 7.98 1H).

IR V (KBr) cm- 1 3420, 1748, 1712, 1681.

Step C-4. Removal of a formyl group To a solution of (2S,4S)-1-t-butoxycarbonyl- 4-formyloxy-2-methoxycarbonylpyrrolidine (5.12 g: 18.7 mmole) in methanol (51.0 ml), aqueous 1N-sodium hydroxide (18.7 ml) is added under ice cooling.

The mixture is stirred at the same temperature for 20 minutes to give (2S,4S)-l-t-butoxycarbonyl- 4-hydroxy-2-methoxycarbonylpyrrolidine 09 g).

Yield: 89%. Colorless crystals.

V0660NMR (CDCl 3 ppm: 1.44(d, J=8.2Hz, 9H), 2.0 to 0Note2.2(m, 1HI), 2.2 to 2.5(m, 1lf), 3.2 to 3.8(m, 3H), *Q 3.79(d, 3=2.8Hz, 3H), 4.2 to 4.5(m, 2H).

IR Y) cm~- 3460, 1728, 1677.

Step C-5. Preparation of a mesyl compound ~In a manner similar to Step A-3 in Prepa*-ative Example 7-A, (2S, 45)-1-t-butoxycarbonyl-4-hydroxy- 2-methoxycarbonylpyrrolidine in dichioromethane is mesylated with triethylamine and methanesulfonylchloo ride in a nitrogen atmosphere under ice cooling to give (2S,4S )-1-t-butoxycarbonyl-4-methanesulfonyloxy-2- 0* *~meth-oxycarbonylpyrrolidine. mp. 90.0 to 91.5'C.

Step C-6. Preparation of a methylol compound In a similar manner to that in Step A-4 in Production Example 7-A, (2S,4S)-l-t-butoxycarbonyl--4methanesulfonyloxy-2-methoxycarbonylpyrrolidinc is 75 allowed to react to give (2S,4S)-l-t-butoxycarbonyl-4methanesulfonyloxypyrrolidine-2-methanol.

Step C-7. Preparation of an acetylthio compound (2S,45 )-l-t-butoxycarbonyl-4-methanesulfonyl oxypyrrolidine-2-methanol a substrate) is allowed to react in a similar manner to Step A-5 in Preparative Example 7-A under a condition for Step C-7 shown in Table 1 to give (2S,4R)-4-acetylthio-.-tbutoxycarbonylpyrrolidine-2.methanol.

NMR 8 (CDCl 3 ppm: 1.47(s, 9H), 2.05 2H), 2.34 3H), 3.0 to 3.3 (in, lx), 3.40(dd, J=ll.6Hz, J=5.2Hz, 1H), 3.5 to 3.9(m, 3H), 3.9 to 4.2(m, 2H).

Step C-8. Introduction of a sulfamide group N-t-butoxycarbonylsulfamide is prepared in the same manner as in the paragraph in Step A-6 in Preparative Example 7-A. (2S,4R)-4-Acetylthio-l-t- *oatbutoxycarbonylpyrrolidine-2-methanol a sub- 20 strate) is allowed to react with N-t-butoxycarbonylsulfamide in the similar manner as in the paragraph in Step A-6 in Production Example 7-A under a condition for Step C-8 shown in Table 2 to give (2S,4R)-4-acetylthio-l-t-butoxycarbonyl-2-.( N-t-butox~vcarbonyl-N- 25 sulfamoylamino) methylpyrrolidinc-.

N NMR 6 (ODd 3 pm 1.41(s, 9H), 1.55(s, 9H), 1.9 to (in, 2H), 2.35(s, 3H), 3.32(dl, J=11.4Hz, J=8.2Hz, 1H), 3.6 to 3.9(m, 3H), 3.9 to 4.1(m, 1H), 4.5(m. lH), 6.15(,s, 2H-).

IR (KBr) cm- 1 3420, 3320, 1706, 1686, 1666.

r r 76 Step C-9. Removal of an acetyl group (2S, 4R) -4-acetylthio-l -t-butoxycarboniyl-2-(Nt-butoxycarbonyl-N-sulfamoylamino) methylpyrrolidine a substrate) is allowed to react in the similar manner as in Step A-7 in Preparative Example 7-A under a condition for Step C-9 shown in Table 3 to give (2S,4R)-l-t-butoxycarbonyl-2--(N-t-butoxycarbonyl-Nsulfamoylamino) methyl-4-mercaptopyrrolidine. mp. 90.0 to 91.5'C.

NMR 6 (CDCl 3 ppm: 1.43(s, 9H), 1.52(s, 9H1), 1.72(d, 1H), 1.9 to 2.0(m, 2H), 3.2 to 514), (in, 1H), 6.11(s, 2H).

IR i)(KBr) cm- 1 3220, 1698, 1683.

Prepa#1 ive Example 7-D of a pyrrolidine derivative 2 HO...O YN--X)OH Y' COH VT -IC *H Boo Boo HO.. tOO 4 lso... AS, N -,CC6 N Boc Boo Boo AcS6 A Boo Bo L*N N,50 2

NH

2

N-'NSO

2

NH

2 Step D-1. Preparation of an N-Boo compound To a suspension of trans -4 -hydroxy- L-prol ine g: 0.381 mole) in methanol (250 ml), a solution of 4N-sodium hydroxide (95.4 ml: 0.381 mole) and di-t- ?7 butyl dicarbonate (91.6 g: 0.42 mole) in methanol ml) is added at -200C. The mixture is stirred at 0 C for 3 hours. The reaction mixture is concentrated and then diluted with toluene (100 ml) and shaken. The aqueous layer is taken, and mixed with conc. hydrochloric acid (36 ml) under ice cooling, saturated brine (100 ml), and ethyl acetate (800 ml). The organic layer is taken, washed with saturated brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue is recrystallized from a toluene-ethyl acetate mixture to give (2S,4R)-l-t-butoxycarbonyl-2-carboxy-4-hydroxypyrrolidine (84.7 g).

Yield: 96%. Colorless crystals. mp. 126 to 128*C.

NMR 6 (CDC1 3 ppm: 1.43, 1.4b(2 x s, 9H), 1.95 to 2.36(m, 2H), 3.36 to 3.6(m, 2H), 4.23 to 4.44(m, 2H).

IR (CHC1 3 cm- 1 3360, 1735, 1656.

eq** Step D-2. Protection of a carboxyl group To a solution of (2S,4R)-l-t-butoxycarbonyl- 4-hydroxy-L-proiine (84.5 g: 0.365 mole) in dichloromethane (1.27 liter) in a nitrogen atmosphere at -30°C, triethylamine (61.1 ml: 0.438 mole) and ethyl chloroformate (38.4 ml: 0.402 mole) are added, and the mixture is stirred for 40 minutes.

Step D-3. Preparation of an O-mesyl compound The resulting reaction mixture containing (2S,4R)-l-t-butoxycarbonyl-2-ethoxycarbonyloxycarbonyl- 4-hydroxypyrrolidine obtained in Step D-2 is cooled to 4 4 -40°C, triethylamine (61.1 ml: 0.438 mole) and methanesulfonyl chloride (31.1 ml: 0.402 mole) are added thereto, and the mixture is stirred for 40 minutes.

78 Step D-4. Reduction To the resulting reaction mixture containing (2S,4R)-l-t-butoxycarbonyl-2-ethoxycarbonyloxycarbonyl- 4-methanesulfonyloxypyrrolidine obtained in Step D-3 cooling at -40°C, tetra-n-butylammonium bromide (11.8 g: 0.0365 mole) and a solution of sodium borohydride (52.5 g: 1.35 mole) in water (55 ml) are added.

The mixture is allowed to warm to -10IC and stirred for 1 hour. The aqueous layer is acidified with dilute hydrochloric acid to pH 3. The organic layer is taken, successively washed with aqueous sodium hydrogen carbonate and water, dried over magnesium sulfate, and concentrated under reduced pressure. The residue is recrystallized from a toluene-hexane mixture to give (2S,4R)-l-t-butoxycarbonyl-4-methanesulfonyloxypyrrolidite-2-methanol (161.3 Yield: 94%. Colorless crystals. mp. 95 to 96 0

C.

NMR 6 (CDC1 3 ppm: 1.48(s, 9H), 1.78 to 2.02(m, 1H), 2.3 to 2.48(m, 1H), 3.05 3H), 3.5 to 3.65(m, 2H), 20 3.65 to 4.0(m, 2H), 4.03 to 4.25 1H), 5.2(s, 1H).

I IR (CHC1 3 cm- 3460, 1680.

Step D-5. Substitution for an acetylthio group A solution of (2R,4S)-l-t-butoxycarbonyl-4- S 25 methanesulfonyloxypyrrolidine-2-methanol a sub- *0 strate) (11.8 g: 40 mmole) and potassium thioacetate (5.94 g: 52 mmole) in dimethylformamide (120 ml) is stirred at 650C for 3.75 hours. The reaction mixture is mixed with ethyl acetate (330 ml), ice water (100 ml), and IN-hydrochloric acid (20 ml) to adjust the aqueous layer at pH 4. The organic layer is taken, successively washed with water and saturated brine, dried over sodium sulfate, and concentrated under I 79 reduced pressure. The residue is purified by silica gel chromatography (toluene-ethyl acetate 2:1) to give (2S,4S)-4-acetylthio-l-t-butoxycarbonylpyrrolidine-2-methanol (9.48 Yield: 86%. Pale orange colored oil.

NMR 6 (CDC1 3 ppm 1.47(s, 9H), 2.34(s, 3H), 2.4 to 3.2(m, 2H), 3.58 to 4.1(m, 6H).

IR (CHC1 3 cm- 1 3380, 1690.

Step D-6. Introduction of a sulfamide group N-t-butoxycarbonylsulfamide is prepared in the same manner as in the paragraph of Step A-6 in Preparative Example 7-A. To a solution of (2S,4S)-4acetylthio-1-t-butoxycarbonylpyrrolidine,2-methanol a substrate) (9.04 g: 32.8 mmole) in tetrahydrofuran (THF) (95 ml), triphenylphosphine (PPh 3 (10.16 g: 38.7 mmole), N-t-butoxycarbonylsulfamide I" (BSMD) (9.66 g: 49.2 mmole), and azodicarboxylic acid diethyl ester (DEAD) (6.20 ml: 39.4 mmole) are succes- 20 sively added under ice cooling. The conditions for this reaction are shown in Table 2, Step D-6. The reaction mixture is diluted with toluene (30 ml), concentrated, diluted with toluene (60 ml), and the formed crystals are filtered off. The filtrate is S 25 concentrated.

Step D-7. Removal of an acetyl group The residue obtained in Step D-6 is dissolved in toluene (95 ml), then, 4.92M sodium methoxide in methanol (20 ml: 98.4 mmole) is added at -35 0 C, and the mixture is stirred for 30 minutes. The reaction mixture is diluted with water (100 ml). The aqueous layer is taken, ethyl acetate (300 ml) is added, mixed with 80 concentrated hydrochloric acid (10 ml) under ice cooling, and the mixture is stirred. The organic layer is taken, successively washed with water and brine, dried over sodium sulfate, and concentrated under reduced pressure. 'The residue is purified by silica gel chromatography. Obtained colorless oil is recrystallized from toluene-hexane mixture to give (2S,4S)-l-tbutoxycarbonyl. 2- N-t -butoxycarbonyl sul1famoyl amino) methyl-4-mercaptopyrrolidine (9.32 Yield: 69%.

Colorless crystals. xnp. 92 to 93 0

C.

NMR (ODC1 3 PPM: 1.2 to 1.5(m, 1H), 1.42(s, 9H), 1..54(s, 9H), 1.82 J=6.2Hz, 1H), 2.5 to 2.7(m, IR), 4.09, 3.05(ABX, J=12.OHz, J=7.4Hz, J=8.2Hz, 2H), 4.06, 3.62(ABX, J=15.OHZ, J=10.8Hz, J=3.2Hz, 2H), 4.2 to 4.6(m, 1H), 6.08(s, 2H).

IR (CHCl 3 cmv-1: 3380, 3220, 1718, 1680.

Elemental Analysis (C 15

H

29

N

3 0 6

S

2 Calcd. C, 43.78; H, 7.10; N, 10.21; S, 15.58 20 Found C, 43.64; H, 7.10; N, 10.19; S, 15.34 81 Table 1 KSAc equiv. a)

DMF

Temp.

o c Time min.

Yield 0 Step C- 7 D-5 1.55 1.20 1.30 1.30 20 fold 5 fold 10 fold 65 10 fold 65 300 105 225 .4 ht 4.

44*4 a.

4 4 4 4 *4*b 4 4 a..

*0

*U~

a) Molar ratio to the substrate b) The volume (ml) of the solvent to the weight of the substrate 0 4 a a 82 Table 2

THF

Stepvo.c PPh 3 equiv. d)

BSMD

equiv. d)

DEAD

equiv. d) Temp. Time min.

Yield A-6 20 fold 1.34 B-6 7 fold 1.50 C-8 10 fold 1.28 D-6 11. fold 1.18 1.20 1. 66 1.50 1. 50 1.30 1.50 45*C 150 76 0 0 C 300 84 1.30 room temp.

1.20 room temp.

240 82 180 -e P a ov a.

4 40* to..

Ge.

of.

The volume (ml) of the solvent to the weight of the substrate Molar ratio to the substrate Not measured 83 Table 3 Na0Me Solventg) Temp. Time Yield Step equiv.f) volih) cmin.

A-7 1.5 15 fold -40 120 72 B-7 2.0 5 fold -10 60 C-9 3.0 4 fold -35 30 D-7 3.0 11 fold -35 30 69 f) Molar ratio to the substrate g) Dichioromethane is used in Steps A-7, B-7, C-9 and toluen~e is used in Step D-7.

h) The volume (ml) of the solvent to the weight of the substrate

U..

4 4* U V a

V

U.

*VVV

S

0 6 a..a

A

84 Prpaaativ ExiaaE!2e rr I dine l~q:o~,~iedervativec lISO. I2 lis,3 ts0.. qI-0 1s02 F 8O 502 Nc~vt Alz Alz C 6

H

4 C1-p Alz AcS*- 4r 'Lc* N NFt I N' N 2 NH 2 Alz Alz Alz Step 1. Preparation of a p-chlorobenzenesulfonyl compound 4*40-alyoyabnl To a solution of (2S,4R)-1-allyloxycarbonyllt** 5 4-methanesulfonyloxypyrrolidine-2-methanol (13.4 g: 6 50 mmole) in dichloromethane (50 ml), r*e tP, p-chlorobenzenesulfonyl chloride (12.66 g: 60 mmole) is added in a nitrogen atmosphere at room temperature and a solution of triethylamine (8.69 ml: 62.5 mmole) in 4S#* 10 dichloromethane (10 ml) is further added dropwise. The mixture is stirred at room temperature overnight. The reaction mixture is successively washed with aqueous sodium hydrogen carbonate and saturated brine, dried over magnesium sulfate, concentrated in vacuo, and 15 purified by silica gel chromatography (toluene-ethyl *P-j acetate) to give crude (2S,4R)-1-allyloxycart, t!- 2 -pchlorobenzenesulfonyloxymethyl- 4 -methanesulfonyloxypyrrolidine (23.73 g) as oil. Yield: 105% NMR 6 (CDC1 3 ppm: 2.2 to 2.6(m, 2H), 3.04(s, 31), 3.58(dd, J=5.0Hz,J=11.4Hz, 1H), 3.8 to 4.0(m, 1H), .1 I I 85 to 4.3(m, 3H), 4.5(m, 3H), 5.1 to 5.4(m, 3H), 5.7 to 1H).

Step 2. Preparation of a phthalimide compound To a solution of (2S,4R)-l-allyloxycarbonyl- 2-p-chlorobenzenesulfonyloxymethyl-4-methanesulfonyloxypyrrolidine (23.7 g: ca. 50 mmole) in dimethylformamide (50 ml), potassium phthalimide (10.2 g: mmole) is added in a nitrogen atmoshpere, and the mixture is stirred at 60°C for 3.5 hours. The reaction mixture is poured into a stirring mixture of ice water (500ml) and ethyl acetate (500 ml). The organic layer is successively washed with water (4 times) and saturated brine, dried over magnesium sulfate, and concentrated in vacuo. The Xesidue is recrystallized from a mixture of n-hexane and toluene. The solid is filtered off and the filtrate is purified by silica ge3 chromatography (toluene-ethyl acetate) to give crude (2S,4R)- S. l-allyloxycarbonyl-2-phthalimidomethyl-4-methanesulfo- 20 nyloxypyrrolidine (12.41 Yield: 61%. Colorless oil.

Step 3. Preparation of an acetylthio compound A solution of (2S,4R)-l-allyloxycarbonyl-2- 25 phthalimidomethyl-4-methanesulfonyloxypyrrolidine '00* (12.4 g: 30.46 mmole) and 90% potassium thioacetate (5.22 g: 45.69 mmole) in dimethylformamide (130 ml) is heated with stirring at 60°C for 4 hours. The reaction mixture is diluted with ethyl acetate (200 ml) and ice water (200 ml). The organic layer is taken, successively washed with water (3 times) and saturated brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel L~L~-P I--I 86 chromatography to give crude (2S,4S)-4-acetylthio-1allyloxycarbonyl-2-phthalimidomethylpyrrol idine (9.33 Yield: 81%.

NMR (C~DC 3 PPM: 1.7 to 1.9(m, I 2.33(s, 314), 2.4 to 2.7(m, 3.25 (dd, J=6.8I1z, J=11. 4Hz,1JH), 3.7 to 4.0(m, 2H), 4.0 to 4.2(m, 2H), 4.3 to 4.6(m, 3H), to 5.3(m, 2H), 5.7 to 5.9(m, 1H), 7.7(m, 2H), 7.85(m, 2H).

Step 4. Removal of a phthalyl and an acetyl groups To a solution of crude (2S,4S)-1-acetylthio- 1-allyloxycarbonyl-2-phthalimidomethylpyrrolidine (5.61 g: 14.90 mmole) in dichioromethane (5.4 ml), and methanol (5.4 ml), hydrazine monohydrate (2.17 ml: 44.7 mmole) is added. The mixture is heated at with stirring for 4 hours. The solid in the reaction mixture is filtered off, washed with dichloromethane ml) and the washing is combined with the filtrate.

The mixture is concentrated to give crude (2S,4S)-2aminomethyl-l-allyloxycarbonyl-4-mercaptopyrrolidine (2.80 Yield: 68%. Oil.

Step 5. Preparation of a sulfamoyl compound To a solution of (2S,4S)-2-.aminomethyl-1allyloxycarbonyl-4-mercaptopyrrolidine (2.80 g: ca.

013.14 mmole) in dichioromethane (66 ml) at triethylamine (4.02 ml: 28.91 mmole) and trimethylchloto to rosilane (3.76 ml: 28.91 mmole) are added dropwise over minutes. The mixture is stirred at the same ternperature for 20 minutes. To the reaction mixture triethylamine (0.92 ml: 6.57 mmole) and a solutian of sulfamoyl chloride (19.37 mmole) in dichioromethane (6.8 ml) are added dropwise over 20 minutes at 87 and the mixture is stirred for 30 minutes. To the reaction mixture triethylamine (3.84 ml: 27.59 mmole) is again added over 1 hour at -500'C. The reaction mixture is kept at the same temperature overnight and concentrated in vacuo. The residual oil is purified by silica gel chromatography (ethyl acetate) to give (2S,4S)-l-allyloxycarbonyl-2-(sulfamoylamino)methyl-4rercaptopyrrolidine (2.64 Yield: 68%. White powder.

NMR '5 (CDCl 3 ppm: 1.4 to 1.6(m, lH), 1.83(d, J=6.2Hz, 1HI), 2.5 to 2.7(m, lH), 3.ll(dd, J=8.2Hz, J-15.2Hz.,lH), 4.13(dd, J=7.3Hz, J=ll.7Hz, 1H), 4.16(dd, J=10.3, 14.9Hz, 1H), 4.3 to 4.6(m, 3H), 4.7(m, 2H), 5.2 to 5.4(m, 4H), 5.8 to 6.0(m, 6.0(m, 2H).

IR 9 (CHCl 3 1 n 1 1684, 1158.

Preparative Example 9 of a pyrrolidine derivative 1 2 j h X10 AIlHC Alz Alz Ste 1.Peaaino nNprtce n n0. S comoun To* a* supnso of(S4lz-ydoy mehxcroylyrl n NhydNochloride (17.0 g: 88 100 mmole) in dichloromethane (200 ml), triethylamine (29.2 ml: 210 mmole) is added in a nitrogen atmosphere under ice cooling. The mixture is stirred for minutes at room temperature, mixed dropwise with a solution of allyl chloroformate (11.2 ml: 100 mmole) in dichloromethane (20 ml), stirred for 1 hour at room temperature, and diluted with water (250 ml). The organic layer is taken, successively washed with water and saturated brine, dried over magnesium sulfate, and concentrated in vacuo to give (2S,4R)-l-allyloxycarbonyl-4-hydroxy-2-methoxycarbonylpyrrolidine (21.82 g) as oil. To a solution of this product in dichloromethane (100 ml), triethylamine (16.7 ml: 120 mmole) and methanesulfonylchloride (9.2 ml: 120 mmole) are added in a nitrogen atmosphere under ice cooling, and the mixture is stirred for 10 minutes.

The reaction mixture is successively washed with aqueous sodium hydrogen carbonate and saturated brine, *oo. dried over magnesium sulfate, concentrated in vacuo, 20 and purified by silica gel chromatography (tolueneethyl acetate) to give (2S,4R)-l-allyloxycarbonyl-4methanesulfonyloxy-2-methoxycarbonylpyrrolidine (27.62 g) as oil. Yield: NMR 6 (CDC13) ppm: 2.2 to 2.4(m, 1H), 2.2 to 25 1H), 2.5 to 2.8(m, 2H), 3.06(s, 3H), 3.74 3.77(2 x s, 3H), 3.8 to 4.0(m, 2H), 4.4 to 4.7(m, 3H), 5.2 to 5.4(m, 3H), 5.8 to 6.0(m, 1H).

S

2. Preparation of a methylol compound To a solution of (2S,4R)-l-allyloxycarbonyl- 4-methanesulfonyloxy-2-methoxycarbonylpyrrolidine (27.12 g: 74.0 mmole) in a mixture of tetrahydrofuran (94 ml) and ethanol (140 ml), sodium borohydride (12 g: 89 31.7 mmole) is added in a nitrogen atmosphere under ice cooling. The mixture is stirred for 4 hours at room temperature. To the reaction mixture concentrated sulfuric acid (8.8 ml: 158.4 mmole) is added dropwise under ice cooling. The reaction mixture is concentrated to half a volum. in vacuo, and diluted with ethyl acetate (100 ml) and ice water (100 ml). The organic layer is taken, successively washed with aqueous sodium hydrogen carbonate and saturated brine, dried over magnesium sulfate, and concentrated in vacuo to give (2S,4R)-l-allyloxycarbonyl-4-methanesulfonyloxypyrrolidine-2-methanol (19.33 Yield: 77 Colorless oil.

NMR 6 (CDCl 3 ppm: 1.9 to 2.1(m, 1H), 2.3 to 1H), 3.05(s, 3H), 3.5 to 3.7(m, 2H), 3.7 to 4.1(m, 2H), 4.1 to 4.3(m, 1H), 4.6(m, 2i), 5.2 to 5.4(m, 3H), 5.8 to 6.1(m, 1H).

osees Step 3. Preparation of an acetylthio compound 20 A solution of (2S,4R)-l-allyloxycarbonyl-4methanesulfonyloxypyrrolidine-2-methanol (19.32 g: 69.17 mmole) and 90% potassium thioacetate (10.73 g: 89.9 mmole) in dimethylformamide (217 ml) is heated with stirring at 65°C for 5 hours. To the reaction 25 mixture ethyl acetate (200 ml) and ice water (200 ml) 0 are added. The organic layer is taken, successively washed with aqueous 0.05N-sodium hydroxide, O.1N-hydro- 0 chloric acid, water and saturated brine, dried over magnesium sulfate, and concentrated in vacuo. The residue is purified by silica gel chromatography to give (2S,4S)-4-acetylthio-l-allyloxycarbonylpyrrolidine-2-methanol (15.34 Yield: 90 NMR 65 (CDC1 3 PPM: 1.5 to 1.7(m, IH), 2.34(s, 31f), 2.4 to 2.6(m, 1H), 3.J,9(dd, J=8.OHZ,J=1l..5Hz, IH), 3.6 to 3.8(m, 2H), 3.8 to 4.0(m, 4.0 to 4.2(m, 2H), 4.6(m, 2H), 5.2 to 5.4(mn, 5.8 to 6.1(m, 1H).

Step 4. Preparation of a sulfamide compound To a solution of (2S,4S)-4-acetylthio-'Lallyloxycarbonylpyrrolidine- 2-methanol (8.02 g: ca.

mmole) in ethyl acetate (83 ml) under ice cooling, triphenylphosphine (9.44 g: 13.6 minole), N-allyloxycarbonylsulfamide (3.12 g: 15.9 mmole), and azodicarboxylic acid diethylester (5.67 ml: 36 mmole) are successively added. The mixture is stirred under ice cooling for 55 minutes and at room temperature for 4 hours. The reaction mixture is dissolved in toluene ml), concentrated, diluted with toluene (60 ml), filtered to remove separating crystals, and the filgoes trate is concentrated. The residue is purified by silica gel chromatography to give (2S,4Sj)-4-acetylthioso 20 1-allyloxycarbonyl-.2-(N-sulfamoyl-N-allyloxycarbonylamino) methylpyrrolidine (6.74 Yield: 55%. Colorless oil.

NMR '5 (CDCl 3 ppm: 1.5 to 1.7(m, 1H), 2.35(s, 3H), to 2.7(m, 1H), 3.19 (dd, J=6.3Hz, J=ll.5Hz, 1H), 3.68(dd, J=3.8Hz, J=14.5Hz, 1H), 3.9 to 4.3(m, 3H), 4.3 to 4.7(m, 5H), 5.2 to 5.4(m, 4H), 5.8 to 6.1(m, 4H).

Step 5. Removal of an acetyl group To a solution of (2S,4S)-4-acetylthio-lallyloxycarbonyl-2-( N-sulfamoyl-N-allyloxycarbonylamino 'methypyrrolidine (6.70 g: 16.4 mmole)in toluene ml), 4.92 M solution of sodium methoxide in metha- 91 nol (5.0 ml: 24.7 mmole) is added at -30'C. The mixture is stirred for 30 minutes, and diluted with water (55 ml). The aqueous layer is takEci diluted with toluene (50 ml), acidified with concentrated hydrochloric acid (2.3 ml) under ice cooling, and stirred. The organic layer is taken, successively washed with water and saturated brine, dried over magnesium sulfate and concentrated in vacuo. The residual oil is purified by silica gel cbi' .o44-tog raphy (toluene-ethyl acetate) to give (2S,4S)-i i!JN.)xycarbonyl-2-(N-sulfamoyl-N-allyloxycarbonylan,, .nyl-4mercaptopyrrolidine (4.89 Yield: 78%. Colorless oil.

NMFR 5 (CDCl 3 ppm: 1.5 to 1.7(m, 1H), 2.35(s, 311), 2.5 to 2.7(m, 1H), 3.19(dd, J=6.3Hz, J=11.5Hz, l1H), 3-68(dd, J=3-8Hz, J=2L4.5Hz, 1H), 3.9 to 4.3(m, 3H), V*Gas4.3 to 4.7(m, 5H1), 5.2 to 5.4(m, 5.8 to 6.1(m, 00*00 S00040 4H1).

0 IR (CHC' 3 cm- 1 1718, 1684, 1179, 1160.

Examples

SA

g* *N

OX

1 OX He OX' a 92 Example X1 2y R ,R -NS02t<R3 1 H PMIB Boo -NHS0 2 NHi 2 2 H PNB Pnz

-NHSO

2

NH

2 3 H PMB PMZ -N1S 2 NA4 4 H PMB PMZ -NHS0 2

N(P=

H P14B PMZ NIIS0 2

N*M

6 Si~e 3 CHPh 2 Boc lS2H

-WM

2

NHC

2 CH0 7 H PNB PrrZ

-HONC

2

H

8 H PMfB PMz -N A~-Pmz 93 Example

R

1 -NH1S0 2 Nll 2 2 -NHS0 2

NH

2 4 4 'WS02NHf -NHSO -M 2 ,fe 6 'N11S0 2 N11 2 7 -NIIS02NHIICH 2 lioH .4 9 -NA 259 94 Example 1. Synthesis of a carbapenem derivative Step Preparation of a protected pyrrolidyithiocarbapenem derivative To a solution of (lR,5S,6S)-6-[(lR)-lhydroxyethyl] -2-oxo-l-methyl-l-carbapenam-3-carboxylic acid p-methoxybenzylester (1.45 g) in acetonitrile ml) at -25*C, diphenyiphosphoric acid chloride (0.953 ml) and diisopropylethylamine (0.872 ml) are successively added. The mixture is stirred at room temperature for 1 hour.. To this mixture 2-sulfamoylaminomethyl-l-t-butoxycarbonyl-4-mercaptopyrrolidine (1.69 g) and d iisopropyl ethyl amine (0.945 ml) are added under ice cooling, and the mixture is stirred for 22 hours. The reaction mixture is mixed with iN-hydrochloric acid (15 ml) and diluted with ethyl acetate.

The organic layer is taken, washed with water, dried over sodium sulfate and concentrated. The residue is purified by column chromatography over 10% wet silica 20 gel to give (lR,5S,6S)-6--[(lR)-l-hydroxyethyl]-2- S) rolidin-3-yl]thio- l-methyl--1-carba-2-penem-3-carboxylic acid p-methoxybenzylester (1.61 Yield: Pale yellow foam.

25 N'-R 6 (CDC1 3 ppm: 1.25(d, J'7.2Hz, 1.32(d, J=6.4Hz, 3H), 1.47(s, 9H), 1.75 to 2.0(m, 1H), 2.4 to 2.65(m, 1H), 2.61(br s, 4H), 3.1 to 3.7 (in, 6F)i, 3.81(s, 3H), 3.75 to 4.25(m, 4H), 5.19, 5.25(A~q, J=12.lIHz, 2H), 6.89(d, J=8.6Hz, 2H), 7. 39(d, J=8.6HZ, 2H-).

IR (CHC1 3 cm- 1 3400, 3290, 1770, 1682.

95 Step 2. Deprotection To a solution of (R,5S, 1ydroxyethyl]-2-[(3S,5S)-5-sulfamoylaminomethyl-l-tbutoxycarbonylpyrrolidin-3-yl]thio-l-methyl-l-carba-2penem-3-carboxylic acid p-methoxybenzylester (1.083 g) in a mixture of dichloromethane (16.5 ml), anisole (1.52 ml) and nitromethane (3.1 ml) at -60*C, a solution of 1.OM aluminum chloride in nitromethane (12.93 ml) is added dropwise. The mixture is stirred for 2 hours slowly warming up to -40°C. The reaction mixture is poured into a solution of sodium acetate (3.18 g) in water (24 ml), successively washed with ether and ether-petroleum ether, desalted and purified by styrene-divinylbenzene copolymer resin column chromatography, and lyophilized the objective fraction to give (1R,5S,6S)-6-[(lR)-l-hydroxyethyl]-2-[(3S,5S)-5sulfamoylaminomethyl-l-pyrrolidin-3-yl]thio-l-methyl-lcarba-2-penem-3-carboxylic acid (0.429 Yield: 67%.

Pale yellow foam.

20 NMR 6 (D 2 0) ppm: 1.22(d, J=7.2Hz, 3H), 1.27(d, J=6.

0 3H), 1.64 to 1.82(m, 1H), 2.62 to 2.80(m, 1H), 3.26 3.59(m, 5H), 3.63 to 3.76(m, 1H), 3.84 to 4.10 2H), 4.16 to 4.29(m, 2H).

0 IR V (KBr) cm- 1 3400, 1750.

25 MIC ml) Staphylococcus aureus strain 3626: 25, Streptococcus pyogenes C203: <0.003.

Example 2. Coupling 2 of a carbapenem derivalive Step 1. Preparation of a Protected pyrrolidylthiocarbapenem derivative To a solution of (1R,5S,6S)-6-[(1R)-1hydroxyethyl]-2-diphenoxyphosphonyloxy-1-methyl-1- 96 carba-2-penem-3 -carboxylic acid p-nitrobenzylester (3.04 g: 5.12 mmole) in acetonitrile (30 ml) under ice cooling, a solution of diisopropylethylamine 16 ml: 1.3 eq.) and 2-sulfamoylaminome-tyl-l-p-nitrobenzyloxycarbonyl-4-xnercaptopyrrolidine (2.4 g: 1.2 eq.) in acetonitrile (20 ml) is added. The mixture is stirred under ice cooling for 140 minutes. The reaction mixture is diluted with ethyl acetate, successively washed with water and saturated brine, dried over magnesium sulfate and concentrated. The residue is purified by silica gel column chromatography (toluene ethyl acetate) to give (lR,5S,6S)-6--[(lR)-l-hydroxyethyl]-2- 5S )-5-sulfamoylaminomethyl-l -p-nitrobenzyloxycarbonylpyrrolidin-3-yl] thio-l-methyl-l-carba-2-penem- 3-carboxylic acid p- nitrobenzyl ester (3.35 g).

Yield: 89P6.

NMER 5 'CDCl 3 ppm: 1.28(d, J=7Hz, 3H), 1.37(d, J=6Hz, got*f 3H), 4.68(s, 2H), 5.22, 5.50(ABq, J=l4Hz, 2H), 5.23(s, 2H), 7.52(d, J=9Hz, 7.65 J=9Hz, 2H), 8.21(d, 20 J=2.5Hz, 2H), 8.26(d, J=2.5Hz, 2H).

IR Q (CHCl 3 cm 1 173 1720, 1704.

p. Step 2. Deprotection To a solution of (lR,5S,6S)-6-[(lR)-lhydroxyethyl]-2-[(3S,5S)-5-sulfamoylaminomethyl-l-pnitrobenzyloxycarbonylpyrrolidin-3-yl] thio-l-methyl-lcarba-2-penem-3-carboxylic acid p-nitrobenzylester (3 g) in a mixture of tetrahydrofuran (60 ml) and 0.lM-MES buffer (pH 10% palladium on carbon (2 g) as a catalyst is added. The mixture is shaken under a stream of hydrogen at atmospheric pressure for 4 hours.

The reaction m,,ixture is filtered to remove the catalyst, washed with ethyl acetate to remove a neutral -97 substance, and concentrated. The residual aqueous solution is purified by styrene-divinylbenzene copolymer resin column chromatography. The fraction eluting with 5 to 10% ethanol water is lyophilized to give (1R,5S,6S)-6-[(1R)-1-hiydroxyethyl]-2-(3S,5s)-5sulfamoylaminomethyl-l-pyrrolidin-3-yl] thio-1-methyl-lcarba-2-penem-3-carboxylic acid (1.42 Yield: 84.8%.

Example 3. Synthesis of a carbapenem derivative Step 1. Preparation of a protected pyrrolidylthiocarbapenem derivative To a solution of (1R,5S,6S)-6-[(lR)-lhydroxyethyl].-2-diphenoxyphosphonyloxy-l-methyl-l- V carba-2-penem-3-carboxylic acid p-methoxybenzylester (1 mmole) in acetonitrile (10 ml) under ice cooling, diisopropylethylamine (1.2 mmole) and 2-p-methoxybenzyloxycarbonylsulfamoylaminomethiyl-1 -p-methoxybenzyloxycarbonyl-4-.mercaptopyrrolidine (1 mmole) are added.

The mixture is allowed to stand overnight. The reaction mixture is diluted with dichloromethane, successively washed with dilute hydrochloric acid, water, aqueous sodium hydrogen carbonate, and 25 brine, dried and concentrated. The residue is purified by silica gel column chromatography to give (lR,5S,6S)-6-[(lR)-l-hydroxyethyl]-2-[(3SS)-sp-methoxybenzyloxycarbonylsui famoylaminomethyl -p-methoxybenzyloxy-carbanylpyrrolidin-3-yl] thio-1--methyl-lcarba-2-penem-3-carboxylic acid p-methoxybenzylester.

Yield :50 to NMR 6 (CDCl 3 ppm: 1.20(d, J=6.4Hz, 3H), 1.34(d, J=6.l-z, 3H4), 3.79(s,9H), 5.00 to 5.12(m, 4H), 5.23, 98 5.15(ABq, J=14.OHz, 2H).

IR (CHC1 3 cm- 1 3390, 1770, 1740, 1693, 1610.

Step 2. Deprotection To a solution Of (lR,5S,6S)-6-[(lR)-1hydroxyethyl3-2- sulfamoylaminomethyl-l1-p-methoxybenzyloxycarbonylpyrrolidin-3-yl] thio-l-methyl-1-carba-2-penem-3carboxylic. acid p-methoxybenzylester (1 rnmole) in dichioromethane (20 mil) at -40 0 C, anisole, (10 mmole) and a solution of 2M aluminum chloride in nitromethane (3 to 4 ml) are added. The mixture is stirred at the same temperature for 1 to 1. 5 hours. The reaction mixture is poured into a solution of sodium acetate (19 to 25 mmole) in water (100 mil), washed with dichlo- VO." romethane to remove a neutral substance. The aqueous %as- layer is purified by styrene-divinylbenzene copolymer resin column chromatography. The objective eluate, is lyophlized to give (lR,5S,6S)-6-((lR)-,l-hydroxyethylj- 20 2 3 S,5S)-5-sulfamoylaminomethyl-lpyrrolidin3.yl]p thio-l-methyl-l-carba-2-penem-3-carboxylic acid.

Yield: 60 to Example 4.__Synthesis of a (3S,5S)-p rrolidylthio- 25 carbapenem derivative Step 1. Preparation of a protected_]pyrrolidylthiacarbapenem derivative To a solution of (lR,5S,6S)-6-[(lR)-1i SIP hydroxyethyl] -2-diphenoxyphosphonyloxy-l-methylcarba-2-penem-3-carboxylic acid p-methox<ybenzylester (700 mg) in acetonitrile (4 nil) at -30'C, a solution of diisopropylethylamine (182 p1) and 1-p-methoxybenzyloxycarbonylN4-mercapto-2- (N-p-methoxybenzyloxycarbo- 99 nyl-N-methylaminosulfonylaminomethyl )pyrrolidine (401 mg) in acetonitrile (3 ml) is added. The mixture is stirred under ice cooling for 90 minutes. The reaction mixture is poured into a mixture of ethyl acetate and dilute hydrochloric acid. The ethyl acetate layer is taken, successively washed with water, aqueous sodium hydrogen carbonate, and brine, dried over magnesium sul-Fit*,, and concentrated in vacuo. The residue is purif, 0 1- silica gel column chromatography (toluene :eth~j I ate =1 to give (1R,.5S,6S)- 6-[(1R)-l-hydroxyetniyl1-2-[(3S,5S)-l-p-meth-oxybo-nzyl- (N -p-methoxybenzyloxycarbo nyl methyl aminosulfonylaminomethiyl )pyrrolidin-3-yl] thio-l-methyl-1-carba-2-penem-3-carboxylic acid p-methoxybenzylester (512 mg).

NMR 6 (CDCl 3 ppm: 1.22(d, 3=7.0Hz, 3H), 1.34(d, J=6.4Hz, 1.6 to 1.9(m, lH), 2.25 to 2.5(m, 1H), to 3.6(m, 7H), 3.778(s, 3H), 3.783(s, 3.788(s, Voo 3H), 5.05(s, 2H), 5.13(s, 2H), 5.2(ABq, 3=12Hz, 2H), 20 6.3 to 6.5(m, 1H), 6.8 to 7.0( m, 6H), 7.2 to 7.4(m, 6H-).

IR 9 (CHCl 3 cm- 1767, 1697.

9* 6 Step 2. Deprotection To a solution of (1R,5S,6S)-6-[(lR--1hydroxyethylj 5S )-l-p-methoxybenzyloxycarbonylp-methoxybenzyloxycarbonyl-N-methylamiiiosul fony\laminomethyl )pyrrolidin-3-yl] thio-l-methyl--carE-Li-2of penem-3-carboxylic acid p-methoxybenzylester (6C mg) 30 in a mixture of dichioromethane (6 ml), i:.trone ane (2 ml) and anis -l (4 ml) stirring at 2-Vsolution of aluminum chloride in nitromethane (2.6 ml: equivalents) is added. The mixture is stirred at 100 ±5 0 C for 1 hour and 30 minutes. The reaction mixture is poured into a mixture of sodium acetate(1.34 water (20 ml) and dichloromethane ml). The aqueous layer is taken, subjected to a styrene-divinylbenzene copolymer resin column chromatography, and the fraction eluting with 8t ethanol is lyophilized to give (1R,5S,6S)-6-[(lR)-1-hydroxyethyl]- 2 3 pyrldi--,]holmty--ab--ee--abx ylic acid (206 mg). Yield 68.6%.

NMR (CDCl 3 PPM: 1.22(d, 3=7.0Hz, 3H), 1.27(d, 3=6.4Hz, 3H), 1.5 to 1.8(m, 1H), 2.63(s, 3H), 2.6 to 2.8(m, 1H), 3.1 to 3.6(m, 5H), 3.65, 3.72(dd, J=6.6HZ, 3=7.6Hz, 1H), 3.8 to 4.4(m, 4H).

IR 2(CHCl 3 cm- 1 1750, 1585.

MIC /ml) Staphylococcus aureus 'n 3626: Streptococcus pyoge nes C203: <0.003.

Example 5. -Synthesis of a 20 carbapenem derivative Step 1. Prepgaration of a protected pyrrolidylthiocarbapenem derivative To a solution of (lR,5S,6S)-6-[(lR)-Ihydroxyethyl]-2-diphenoxyphosphonyloxylmeth.l-lcarba-2-penem-3.-carboxylic acid p-methoxybenzylester mmole) in acetonitrile (10 ml) under ice cooling, di-isopropylethylamine (1.2 mmole) and 2-N, N-dimethylsulfamoylaminomethyl-1 -p-methoxybenzyloxycarbonyl-4- .*foe:mercaptopyrrolidine (1 mmole) are added. The mixture is allowed to stand overnight. The reaction mixture is diluted with dichloromethane, successively washed with dilute hydrochloric acid and water, dried, and concentrated. The residue is purified by silica gel column 101 chromatography to give (lR, 5S, 1R)-1-hydroxyethyll 3S, 5S N-dimethylsulfamoylamii'omethyl-1-pmethoxybenzyloxycarbonylpyrrolidin-3-yl] thio-1-methyl- 1-carba-2-penem-3-carboxylic acid p-methoxybenzylester.

Yield: 50 to NMR 5 (CDCl 3 ppm: 1.22(d, J=7.2Hz, 3H), 1.34(d, J=6.2Hz, 3H), 2.76(S, 6H), 3.79(s, 3H), 3.81(s, 3H), 5.06(s, 2H), 5.24, 5.18(ABq, J=l2Hz, 2H).

IR V) (CHC1 3 cm- 1 3390, 1770, 1725, 1690, 1610.

Step 2. Deprotection A solution of (lR,5S,6S)-6--[(lR)-1-hydroxyethyl]-2-[ (3S,5S)-5-N,N-dimethylsulfamoylaminomethyl-lp-methoxybenzyjoxycarbonylpyrrolidin-3 -yl] thio- 1- .methyl-l-carba-2--penem-3-carboxylic acid p-methoxybenzylester (1 mmole) in dichloromethane (20 ml) is *cooled to -40*C. Anisole (10 mmole) and a solution of 2M aluminum chloride in nitromethane (3 to 4 ml) are 0. ~~**added thereto, and the mixture is stirred at the same 20 temperature for 1 to 1.5 hours. The reaction mixture is poured into a solution of sodium acetate (19 to mmole) in water (100 ml), and washed with dichloroo *methane to remove a neutral material. The aqueous layer is purified by styrene-divinylbenzene copolymer *25 resin column chromatography and the objective eluate is lyophilized to give (lR,5S,6S)-6-[(1R)-1-hydroxyethyl]- 2- 5 N-dimethylsulfamoylaminomethylpyrro- 0* 0 lidin-3-yl] thio-l-methyl-l-carba--2-penem-3-carboxylic acid. Yield: 60 to NMR 5 (D 2 0) ppm: 1.2(d, J=7.4Hz, 1.28(d, 3=6.4Hz, 3H), 1.65 to 1.80(m, 1E), 2.65 to 2.80(m, 1H), 2.81(s, 6H), 3.29 to 3.55(m, 5H), 3.65 to 3.75(m, 1H), 3.80 to 4.10(m, 2H), 4.16 to 4.30(m, 2H).

d -102 IFR I) (KBr) cuif': 3400, 1750.

MIC Lr ml) :Staphylococcus aureus strain 3626: Streptococcus pyogenes 0203: <0.003.

Example 6. Synthesis of a carbapenem derivative Step 1. Preparation of a protected pyrrolidylthiocarbapenem derivative To a solution of (2R,5S,6S)-2-diphenoxyphosphonyloxy-6- lR)-l-hydroxyethyl] -l-methyl-l-carba- 2 -penem-3-carboxylic acid diphenylmethylester (6.88 g: 11 mmole) in dichloromethane (70 ml) under ice cooling, trimethylchlorosilane (1.81 ml: 14.3 mmole) and triethylamine (1-.99 ml: 14.3 mmoie) are added. The mixture is stirred for 25 minutes. The reaction mix- V9696ture is poured into aqueous sodium hydrogen carbonate.

The organic layer is taken, washed with water and brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue containing the product, (lR, 5S, 6 S)-2-diphenoxyphosphonyloxy--methyl.

6- lR)-l-trimethylsilyloxyethyl] -l-carba-2-penem-3carboxylic acid diphenylmethylester is dissolved in acetonitrile (70 ml), and (2S.4S)-l-t-butoxycarbonyl- 2 -(N-t-butoxycarbonyl-N-sulfamoylamino )methyl-4-mercaptopyrrolidine (5.43 g: 13.2 mmole) and diisopw,:pvlethylamine (2.30 g: 13.2 mmole) are added thereto under ice cooling. The obtained mixture is stirred for hours. To the reaction mixture containing the a a product, (lR,5S, 6 S)--2-[(3S,5S)-l-t-butoxycarb-nyl-5-(Nt-butoxycarbonyl-N-sulfamoylanlino)methylpyr- 11ifdin-3carba-2-penem--3-carboxylic acid diphenylmet'nylestar, iN-hydrochloric acid (5.5 ml) is added, and the mixture 103 is stirred for 20 minutes, diluted with ethyl acetate(150 ml), and the mixture is poured into ice water. The organic layer is taken, successively washed with aqueous sodium hydrogen carbonate, water, and brine, dried over sodium sulfate, and concentrated under reduced pressure. The residue is recrystallized from toluene to give (lR,5S,6S)-2-jI(3S,5S)-l-t-butoxy- (N-t-butoxycarbonyl-N-sulfamoylamino )methylpyrrolidin-3-yl] thio-6- lR) -l-hydroxyethyl) -1-methyll-carba-2-penem--3-carboxylic acid diphenylmethylester (7.53 Yield: 87%. Colorless crystals. mp.163 to 16400.

NMR 6 (CDCl 3 ppm :l.27(d, J=7.2Hz, 3H), 1.39(s, 9H), 1.42(s, 9H), 2.45 to 2.65(m, 1H), 3.1 to 3.35(m, 2H), 3.28(dd, J=7.2Hz, J=2.6Hz, 1H),3.5 to 3.77(m, 2H), 3.9 to 4.15(m, 2H), 4.26(dd, J=7.0Hz, J=2.6Hz, 1H), 4.2 0 to 4.37(m, 1H), 4.45 to 4.66(m, 1H), 6.07(s, 2H), 6.95(s, lH), 7.2 to 7.6(m, IR V (CHCl 3 cm- 1 3385, 3230, 1778, 1715, 1685.

20 Elemental Analysis (C 38 H ON 4 0 0 Calcd.: C, 57.99; H, 6.40; N, 7.12; S, 8.15.

Found 57.87; H, 6.46; N, 6.99; S, 7.93.

Step 2. Deprotection 25 To a solution of aluminum chloride (3.20 g: 24 mmole) in a mixture of anisole (24 ml) and dichioromethane (24 ml) at .4000, a solution of (7R,5S,6S)-2- 99*[(3S,5S)-l-t-butoxycarbonyl-5.-(N-t-butoxycarbonyl-Nsul famoyl amino) methylpyrrolid in- 3-y'L] thio-6-[(lR)-1hydroxy-ethyl] -l-methyl-l-carba-2-penem-3-carboxylic acid diphenylrnethyleste r (2.36 g: 3 mmole) in dichloromethane (12 ml) is dropwise and gradually added. The mixture is vigorously stirred at -25 to -30'C for 104 hours. The reaction mixture is poured into a solutio: of sodium acetate (5.91 g: 72mmole) in water (48 ml). The aqueous layer is taken, washed with dichloromethane, concentrated in vacuo to remove remaining orgaic solvent and subjected to styrene-divinylbenzene copolymer resin column chromatography. The fraction eluting with methanol-water is lyophillized to give (lR,5S,6S)-6-jI(lR)-1-hydroxyethyl]-2- (3S, 5S) -5-sulf amidomethy'Lpyrrolidin-3-yl] thio-1-methyl-l-carba-2-penem-3-carboxylic acid (910 mg) Yield: 72%. Colorless foam.

NMR 8 (D20) ppm: 1.22(d, J=7.2Hz, 3H), 1.27(d, J=6.3Hz, 3H), 1.64 to 1.82(m, 1H), 2.62 to 2.80(m, 1H), 3.26 to 3.59(m, 3.63 to 3.76(m, 3.84 to 4.10(m, 2H), 4.16 to 4.29(m, 2H).

IR cnf 1 3400, 1750.

MIC Iml) taphylococcus aureus 3626: Blood level: mice i a fter 15 min (2r ml): 9.8.

Urinary recovery: mice 36.3.

*se Example 7. Synthesis of a carbapenem derivative o Step 1. Preparation of a protected pyrrolidylthiocarbapenem derivative To a solution of (lR,5S,6S)-6-[(lR)-1hydroxyethyl] -l-methyl-2-oxo-l-carbapenam-3-carboxylic acid p-methoxybenzylester (277 mg) in acetonitrile (4 ml) under ice cooling, diphenylphosphoric acid chloride (198 p1) and diisopropylethylamine (166 p1) are successively added. The mixture is stirred at room temperature for 1 hour. To the reaction mixture containing the product, (iR, 5S, 6S)-2-diphenoxyphosphionyloxy-6-[(lR)-1-hydroxyethyl]-l-methyl-l-carba-2-penemi~- 3-carboxylic acid p-methoxybenzylester, (2S,4S)-2-(2hydroxyethyl)sulfamoylaminomethyl-l-p-methoxybenzyloxy-carbonyl-4-mercaptopyrrolidine (344 mg) and diisopropylethylamine (166 pl) are added under ice cooling, and the mixture is stirred at the same temperature for 2 hours. The reaction mixture is diluted with ethyl acetate, successively washed with water, dilute hydrochloric acid, water, aqueous sodium hydrogen carbonate and water, dried over magnesium sulfate, and concentrated. The residue is purified by silica gel column chromatography to give (1R,5S,6S)-6-[(lR)l-hydroxyethyl]-2-[(3S,5S)-5-(2-hydroxyethyl)sulfamoylaminomethyl-l-p-methoxybenzyloxycarbonylpyrrolidin-3yl]thio-l-methyl-l-carba-2-penem-3-carboxylic acid p-methoxybenzylester (156 mg). Yield: 26%.

NMR 6 (CDC1 3 ppm: 1.22(d, J=7.0Hz, 3H), 1.34(d, J=6.2Hz, 3H), 3.79(s, 3H), 3.80(s, 3H), 5.05(s, 2H), 0* o o 5.17, 5.24(ABq, J=12.2Hz, 2H).

IR V (CHC1) cm 1 1775, 1690.

S 00 Step 2. Deprotection To a solution of (lR,5S,6S)-6-[(1R)-1hydroxyethyl]-2-[(3S, 5S)-5-(2-hydroxyethyl)sulfamoylaminomethyl-l-p-methoxybenzyloxycarbonylpyrrolidin-3- 25 yl]thio-1-methyl-l-carba-2-penem-3-carboxylic acid p-methoxybenzyl ester (148 mg) in a mixture of dichloromethane (3 ml) and nitromethane (750 pl) in a nitrogen atmosphere at -40°C, a solution of 1.OM aluminum 09 chloride in nitromethane (1.8 ml) and anisole (258 pl) 0 is added. The mixture is stirred at the same temperature for 1.5 hours. The reaction mixture is poured into a solution of sodium acetate (454 mg) in water (8 ml) and washed with an ether-hexane mixture. The

I

106 aqueous layer is concentrated in vacua to 4 ml, and purified by styrene-divinylbenzene copolymer resin column chromatography to give (lR,5S,69)-6-[(lR)-lhydroxyethylj.-2-[(3S,5S)-5-(2-hydroxyethyl)sulfamoylaminomethylpyrrolidin-3-ylJ thio-l-methyl-l-carba-2penem-3-carboxylic acid (42 mg). Yield: 46%.

NMR 6 (D 2 0) ppm: 1.21(d, 3=7.4Hz, 3H), 1.28(d, 3=6.4Hz, 3H), 1.66 to 1.81(m, 2.66 to 2.81(m, lH), 3.15(t, 3=5.6Hz, 2H), 3.)2 to 3.54(m, 511), 3.65 to 3.75(m, 311), 3.87 to 4.07(m, 2H), 4.18 to 4.27(m, 2H).

IR (KBr) cm- 1 3400, 1750.

Blood level: mice after 15 min ml): 29.3.

Example 8. Synthesis of a :**Soobapenem derivative .,..Step 1. Preparation of a protected pyrrolidyithiocarbapenem derivative To a solution of (lR,5S,6S)-2-diphenoxyphosphonyloxy-6-[ (lR)-1-hydroxyethyll -l-methyl-l-carba- 2-penem-3-carboxylic acid p-methoxybenzylester (456 mg) in acetonitrile (3 ml) under ice cooling, diisopropy- :lethylamine (165 -pl) and (2S,4S)-2.-(l,l-dioxo-2-pmethoxybenzyloxycarbonyl-l, 2, 25 methyl-4-mercapto-l-p--methoxybenzyloxycarbonylpyrrolidine (445 mg) are added, rIne mixture is allowed to stand at 000 overnight.. The reaction mixture is diluted, with ethyl acetate, successively washed with water, dilute hydrochloric acid asnd water, dried over magnesiurn sulfate, and concentrated in vacua. The residue is purified by silica gel chromatography to give= (lR,5S,6S)-2-[(3S5S)-5)-(1,1-dioxo-2-p-metboxybenzyl- .oxycarboiiyl-1,2,5-thiadiazolidin-5-yl) methyl-l-p-m~eth- 107 oxybenzyloxycarbonylpyrrolidin-3-yl] thio-6- -1hydroxyethyl] -l-methyl-l-carba-2-penem-3-carboxylic acid p-methoxybenzylester (510 mg). Yield: 72%.

NMR 6 (ODC1 3 ppm: 1.22(d, 3=7.4Hz, 3H), 1.34(d, J=6.2Hz, 3H), 5.04(s, 2H), 5.23(s, 5.18, 5.24(ABq, IR \2 (CHCl 3 cm'1: 1773, 1735, 1700.

Step 2 Deprotection To a solution of (lR,5S,6S)-2-[(3S,5S)-5- (l,l-dioxo-2-p-methoxybenzyloxycarbonyl-1,2, )methyl-l-p-methoxybenzylojxycarbonylpyrrolidin-3-yljthio-6-[ (lR)-l-hydroxyethylj -1-methyll-carba-2-penem-3-carboxylic acid p-methoxybenzylester (500 mg) in~ a mixture of dichloromethane (8 ml) and n.Ltromethane (3 ml) in a nitrogen atm~osphere at -40 0

C,

anisole (729 p1) and a solution (5.03 ml) of l.OM aluminum chloride in nitromethane re added. The mixis stirred at the same temperature for 1.5 hours.

The reaction mixture is poured into a soution of sodium acetate (1.28 g) in water (50 ml), then aqueous layer is taken, and washed with an ether-hexane mixture. The aqueous layer is concentrateC~ under reduced pressure to about 15 ml, and is purified by styrene -d ivinylbenzene copolymer resin column chromatography to give (lR,5S,6S)-2-[(3S,5S)-5-(1,1-dioxo-1,2,5-thiadiazoli- )methylpyrrolidin-3-yl] thio-6- j( lR) -1-hydroxyethyl]-l-methyl- l-carba-2-penem-3-carboxylic acid 4 (180 mg). Yield: 72%.

NMR 5 (D20) ppm: 1.21(d, 3=7.4Hz, 3H), 1.28(d, 3=6.4Hz, 3H), 1.68 to 1.84(m, 1H), 2.71 to 2.85(m, 1H), 3.28 to 3.77(m, 10H), 3.94 to 4.12(m, 2H), .17 to 4.31(m, 2H).

109 Step Deprotection To a solution of (lR,5S,6S)-2-[(3S,5S)-5l-dioxo-2--p-methoxybenzyloxycabolyl-3, 4, 5,6-tetrahydra-i, 2, 6-thiadiazin--6-yl)methyl-l--p-methoxybenzyl.oxycarbonylpyrrolidin-2 -yl~thio-6-(R)-1I-hydroxyethyl] -l-methyl-l-carba-2-penem-3-carboxylic acid p-methoxybenzylester (600 mg) in a mixture of dichioromethane (9 ml) and nitromethane (3.5 ml) in a nitrogen atmosphere at -40*C, anisole (861 p1) and a solution of 1.OM aluminum chloride in nitromethane (5.94 ml) are added. The mixture is stirred at the same temperature for 1.5 hours. The reaction mixture is poured into a solution of sodium acetate (1.52 g) in water (50 ml), and washed with a mixture of ether and hexane. The aqueous layer is concentrated in vacuo to about 15 ml, and the mixture is purified by styrene-divinyibenzene copolymer resin column chromatography to give (lR,5S,6S)-.2-[(3S,5S)-5-(1,1--dioxo--3,4,5,6-tetrahydrofew.* 1,2, 6-thiadiazin--6-yl)methylpyrrolidin-3-yl] thio-6- [(lR)-l-hydroxy--ethyl]-l-methyl-l-carba-2-penem-3carboxylic acid (190 mg). Yield: 63%.

*:rN .MR 6 (D0 pp:1.20(d, J=7.2Hz, 3H), 1.27(d, of* J=6.4Hz, 3H), 1.65 to 1.80(m, 3H), 2.65 to 2.60(m, 1H), 3.27 to 3.56(m, 9H), 3.64 to 3.74(m, IH), 3.91 to 4.10(m, 2H), 4.15 to 4.30(m, 2H).

IR cm-1: 3400, 1750.

oMIC Staphylococcus aureus strain 3G26! 4. w 25. Blood level: mice after 15 min. (4/ml): 28.4.

110 Examples 10 to 12. Synthesis of (3R, 5R), (3R, 5S) and 5R) pyrrolidylthiocarbapenem derivatives 011 Me OH 1V- Boc 1 (OPh)2 2-\No GOOCHPh 2 ODOHPh 2 OH Mie

ODOH

Step 1. Preparation of a protected pyrrolidyithiocarbapenem de:civatives To a solution of (lR,5S,6S)-2-diphenoxy- ~phosphonyloxy-6- R) -1-hydroxyethyl] -l-methyl-l-carba- 2-penem--3-carboxylic acid diphenylmethyl ester a substrate) and 1-t-butoxycarbonyl-2- (N-t-butoxycarbonyl-N--sulfamoylamino )methyl-4-mercaptopyrrolidine (Pyld) in acetonitrile (MeCN) under ice cooling, diisopropylethylamine (HNPr-i) is added dropwise. The mixture is stirrud to react under a condition shown in ~*Table 4. The reaction mixture is diluted with ethyl acetate, and ice water is added thereto. The organic layer is taken, successively washed with water and .~*saturated brine, dried over magnesium sulfate, and Of* 15 concentrated in vacuo. The residue is purified by silica cel chromatography to give (lR,5S,6S)-2-[l-t-- N-t-butoxycarbonyl-N-sulfamoylamino) methylpyrrolidin-3-yl] -thio-6- I R)-l--hydroxyethyl] -1methyl-l-carba-2-penem-3-carboxylic acid diphenylmethyl ester.

ill Table 4 Configuration of Pyldj) HNPr-ij)MeCNk) Temp. Time Yield Example Pyld equiv. equiv. vol. mil. 9 Ex. 10 3R5R 1.17 1.30 7 ice 240 86 fold cooling Ex. 11 3R5S 1.20 1.30 7 ice 120 88 fold cooling Ex. 12 -3S5R 1.14 1.27 7 ice 270 73 fold cooling

U

9* *9 9

S..

S

9.4%' 9**

U..

9. 9 S* 9*S* a.

.a a *9 1) Configuration of a pyrrolidine ring j) Molar ratio to the substrEzte k) The volume (ml) of the solvent to the weight of the substrate 112 Ph ysical properties of the products Isomer: NMR 5 (CDC1 3 ppm: 1.26(d, 3=7.2Hz, 3H), 1.39(d, 3=6.2Hz, 3H), 1.43 9H), 1.51(s, 9H), 2.5(m, 1H), 3.1 to 3.9(m, 6H), 4.0 to 4.7(m, 4H), 6.i.(mn,1H), 6.98(s, 1H), 7.1 to 7.6(m, IR 9 (KBr) cm- 1 3400, 3240, 1770, 1710, 1670.

Isomer: NMR 6 (CDC1 3 ppm: 1.28(d, 3=7.0Hz, 3H), 1.36(s, 9H), 1.40(d, J=6.2 Hz, 3H), 1.52(s, 9H), 2.0(m, 1H), 3.2 to 3.9(m, 7H), 4.2 to 4.4 (in, 2H), 4.4 to 4.6(m, 1H), 6.01(s, 2H), 6.94(s, 1H), 7.1 to 7.6(m, IR V) (KBr) cm- 1 3400, 3240, 1772, 1708, 1682.

!(3R,5S) Isomer: NMR 5 (CDCI 3 ppm: 1.76(d*, 3=7.2Hz, 3H), 1.3 to 1.5(m, 12H), 1.52(s, 9H), 1.9 to 2.1(mn, 1H), 3.2 to 3.9(m, 7H), 4.1 to 4.4(m, 2H), 4.4 to 4.6(m, 1H), 6.04(s, 2H), 6.94(s, 1H), 7.1 to 7.6(m, IR V (KBr) cnf 1 3420, 1770, 1710.

9 :.Step 2. Deprotection A solution of (lR,5S,6S)-2-[1-t-butoxyto o .carbonyl-5- (N-t-butoxycarbonyl-N-sulfamoylamino )methyl- 25 pyrrolidin-3-yllthio-6-[(1R)--1-hydroxyethyl] -1-methyl- 1-carba-2-penem-3--carboxylic acid diphenylmethyl ester to a substrate) in dichioromethane (DCM) is added dropwise in a nitrogen atmosphere into a solution of aluminum chloride (Aid1 3 in a mixture of dichloromethane (DCM) and anisole (PhOMe). The mixture is stirred to react under zhe condition shown in Table 5. To the reaction mixture, ac;*jeous sodium acetate is added. The aqueous layer is taken, washed with dichioromethane, 113 and purified by column chromatography over styrenedivinylbenzene copolymer resin to give (lR,5S,6S)-6- C( lR)-1-hydroxyethyl] 3-yl] thio-1-rnethyl-1-carba-2-penem-3-carboxylic acid.

Table Configuration') AlCl 3 m) DCMn) PhOMen)Temp. Time Yield Example of Pyld equiv. vol. vol. cc min. 0- (fold) (fold) Ex. 10 3R5R 8.0 16 10 -30 300 86 Ex. 11 3R5S 8.0 17 10 -30 150 88 Ex. 12 3S5R 8.0 17 10 -30 150 73 ft ft ft...

ft. ft ft ft ft ft...

ft.

ft ft. ft ft...

ft...

ft.

ft.

1) Configuration of a pyrrolidine ring m) Mo~lar ratio to the substrate n) The volume (ml) of the solvent to the weight of the substrate 114 Physical properties of the products Isomer: NMR 6 (D 2 0) ppm: 1.18(d, J=7.2Hz, 3H), 1.27(d, 3=6.2Hz, 3H), 1.9(m, 1H), 2.7(m, 1H), 3.2 to 3.6(m, 3.6 to 3.8(m, 1H), 3.8 to 4.1(m, 2H), 4.2(m, 2H).

IR V (KBr) cm': 3360, 1750.

Isomer: NMR 6 (CD 3

SOCD

3 ppm: 1.09(d, 3=7.0Hz, 3H), 1.14(d, J=6.2Hz, 3H), 1.7 to 2.O(m, 1H), 1.9 to 2.2(m, 1H), 2.9(m, 1H), 3.0 to 3.3(m, 4H), 3.3 to 3.6(m, 1H), 3.6 to 3.8(m, 2H), 3.9(m, 1H), 4.1(m, 1H).

IR Y (KBr) cm- 1 3340, 1765, 1740, 1620, 1575, 1548.

Isomer: 15 NMR 6 (D 2 0) ppm: 0.86(d, J=7.4Hz, 3H), 0.93(d, J=6 .4Hz, 3H), 2.43(d,J=6.4Hz, 311), 1.90(dd, 3=4.4Hz, 2H), 2.9 to 3.3(m, 5H), 3.48(dd, J=13.2Hz, 3=7.2Hz, 1H), 3.7 to 3.8(m, 21), 3.8 to 4.0 21), 4.47 DHO.

IR y (KBr) cmf 1 3400, 1750, 1585.

Example 13. Synthesis of a bapenem derivative using a monoallyloxycarbonyl intermediate HO M Me3Si0 Me Me 3 SiO be.

Er, O (OPh)2 shKNcosh 1 -LSin$\/-oho( O~ =2CH:CRCH2 GO3OCHZ(TC112 COXC 2 CH=CH2 OH la OH Me 0 NHON2~1S 2 h 3 4 cm CH 2

CW

CDYXHCH=CH2

CXXOH

115 Step 1 .Preparation of a trimethylsilyl compound To a solution of (3S,4S)-3-[(lR)-1-hydroxyethyl]-4-[( R)-l-phenylthiocarbonylethyl]-1-allyloxycarbonylmethyl-2-azetidinone (5.04 g: 13.35 mmole) in toluene (40 ml) under ice cooling, pyridine (1.51 ml: 18.69 mmole) is added and trimethylchlorosilane (2.26 ml: 17.36 mmole) is added dropwise. The mixture is stirred at room temperature for 1.5 hours. Water ml) is added to the reaction mixture, and aqueous layer is extracted with toluene. The extract and the organic layer are combined, washed with water (2 times) and saturated brine, dried over magnesium sulfate, and concentrated in vacuo to give crude trimethylsilyloxyethyl]-4-[(1R)-l-phenylthiocarbonyl- 15 ethyl]-l-allyloxycarbonylmethyl-2-azetidinone (5.614 g) as oily residue. Yield: 94%.

Step 2. Ring closure To a solution of the crude trimethylsilyloxyethyl]-4-[(lR)-l-phenylthiocarbonylethyl]-1-allyloxycarbonylmethyl-2-azetidinone (5.60 g: 12.454 mmole) obtained in Step 1 in tetrahydrofuran (62 ml) at -60°C, a solution of iM-potassium t-butoxide (24.9 mmole) in tetrahydrofuran (24.9 ml) is added *ee* 25 dropwise. The mixture is stirred for 10 minutes.

After adding iodomethane (0.48 ml: 14.94 mmole) and stirring at the same temperature for 20 minutes, diphenylphosphoryl chloride (2.73 ml: 12.45 mmole) is added thereto. After allowing to warm to an ice water temperature over 1 hour, the reaction mixture is diluted with toluene (120 ml) and water (120 ml). The aque -us layer is extracted with toluene. The extract and the organic layer are combined, successively washed with 116 water (2 times), aqueoijs sodium hydrogen carbonate, and saturated brine, dried over magnesium sulfate, and concentrated in vacuo to give crude (lR,5S,6S)-2-diphenoxyphospholyloxy-6- (lR)-l-trimethylsi-lyloxyethyl] -1methyl-carba-2-peflem-3-carboxylic acid allyl ester (3.795 g) as oily residue. Yield: 104%.

IR y (CHC1 3 cm- 1 3008, 1778, 1722, 1636, 1589, 1489.

NMR 6 (CDCl 3 ppm: 0.12(9H, 1.19(3H, d, J=7.2Hz), 1.25(3H, d, J=6.2Hz), 3.24(lH, dd, J=3.O11z, J=6.8Hz), 3.3 to 3.6(1H, in), 4.ll(lH, dd, 3=3.0Hz, 3=10.2Hz), 4.1 to 4.3(lH, in), 4.6 to 4.7(2H, in), 5.1 to 5.5(2H, in), 5.7 to 6.0(lH, mn), 7.1 to 7.5(10H, in).

Step 3. Preparation of a protected pyrrolidyithiocarbapenen derivative *To. a solution of crude (1R,5S,6S)-2phenoxy-phospholyloxy-6-[( 1R)-l-triinethylsilyloxyethyl] methyl-1-carba-2-penein-3-carboxylic acid allyl ester (2.56 g: 4.2 mmole) obtained in Step 2 and 4S allyloxycarbonyl-2- (N-sulfainoylamino )methyl- 5:4-mez-captopyrrolidine (1.4F g: 5.0 mmole) in acetonitri. (13 ml) under ice cooling, diisopropylethylanine ml- 5.46 iniole) is added dropwise, and the mixture stirred at the same temperature for 7.5 hours.

The reaction mixture is acidified with 1N-hydrochloric acid (6.3 ml), stirred at the same temperature for minutes, and ethyl acetate (80 ml) and ice water ail) are added thereto. The organic layer is taken, successively washed with water and saturated brine, dried over magnesium sulfate an cc--.centrated in vacUo.

The residue is purified by silica gel chromatography (tollu;ne-ethyl acetate) to give (lR,5S, 6S)-2-II(3S,5S)- 117 (N-suJfamoylamino)ethyl-pyrrolidii.-3-y]thio-6-[(R)-l-doxhyrxety)--ehy--cr 2-penem-3-carboxylic acid allyl ester (1.63 g).

Yield: 71%.

IR y) (CHCl 3 cm-1: 1772, 1691, 1410.

1 HNMR 5 (CDCl 3 ppm: 1.26(3H, d, J=7.0Hz), 1.35(3H, d, 3=6.0Hz), 1.7 to 2.7(3H, mi), 3.1 to 3.5(5H, to 3.8(1H, 3.9 to 4.4(4H, 4.5 to 4.9 (4H, m), to 5.5(5H, 5.8 to 6.1(2H, m).

Step 4. Deprotection To a solution of (lR,5S,6S)-2-[(3S,5S)-l- N-sulfamoylamino) methylpyrrolidin- 3-yl]thio-6-[(1R )--hydroxyethyl]-l-methyl-l-carba-2- 15 penem-3-carboxylic acid allyl ester (379 mg: 0.695 miole) in acetone (14 ml), triphenylphosphine mg: 0.21 mmole) and tri-n-butyltin hydride (0.424 ml: 1.53 mmole) are added. Under ice cooling palladium tetrakis(triphenylphosphine) (81 mg: 0.07 mmole) is further added. After stirring at the same temperature for 45 minutes and at room temperature for 1 hour, water (35 ml) and methylene chloride ml) are added to the reaction mixture. The aqueous layer is taken, washed with methylene chloride and lyophilized to give (lR,SS,6S)-6-[(lR)-l-hydroxyethyll- 2-[(3S,5S)-S-sulfamidomethylpyrrolidin-3-ylthio-linethyl--carba-2-penem-3-carboxylic acid (238 mg).

Yield: 82%. (HPLC purity NMR 5 (D 2 0) ppm: 1.22(d, 3=7.2Hz, 3H), 1.27(d, 3=6.3Hz, 3H), 1.64 to 1.82(m, 1H), 2.62 to 2.0(m, lH), 3.26 to 3.59(m, 5H), 3.63 to 3.76(i, 1H), 3.84 to 4.10(, 2H), 4.16 to 4.29(m, 2H).

IR (KBr) 1340, 1750.

I p II 118 Example 14. Synthesis of a bapenem derivative using a diallyloxycarbonyl intermediate HO Mie lMe 3 SiO lie_ JEkN<OSPh LjK h CcOCH 2 CI4=CH 2

CXOCHZCH=CH

2 OH lie Alz 01 3 O iNI Alz 4 O

CGOXCH

2

CH=CH

2 lIe3SiO Hie J, PO(OPh) 2 2 0

ET

COOJH

*9e~ 0*9 9*99 9* 99 9 9 9 9 9 9 .9 9O*9 o 9 89 9 0 09 9 Step 1. Preparation of a trimethylsilyl compound 5 (3S,4S)-3-[(lR)..1-hydroxyethyll--4-[(lR)-lphenylthiocarbonylethyl]l1-allyloxycarbonylmethyl -2azetidinone is trimethylsilylated in the same manner as in Step 1 in Example 13 to give trimethylsilyloxyethyl]-4-i:( iR) -l-phenyithiocarbonylethyl Ij-1-allyoxycarbonylrnethyl-2-azetidinone.

Step 2. Ring closure The crude (3S,4S)-3-[(lR)--l-trimethylsilyloxyethyl] (lR)-1-phenylthiocarbonylethyl] -1-ally- 15 loxy-carbonylmethyl-2-azet'idinone obtained in Step 1 is allowed to react to close the ring in the similar manner as in Step 2 of Example 13 to give crude (lR, SS,6S) -2-diphenoxyphosphoryloxy-6- II( R trimethylsilyloxyethyl] -l-methyl-1-carba-2-penem-3-carboxylic acid allyl ester.

119 Step 3 Preparation of a protected pyrrolidyithio carbapenem_,derivative Under similar reaction condition, the crude (lR,5S,6S)-2-diphenoxyphosphoryloxy-6-(1R)-l-trimethylsilyloxyethyl)-l-methyl-l-carba-2-penem-3-carboxylic acid allyl ester (5.05 g: 8.3 mmole) obtained in Step 2 and (2S,4S)-l-allyloxycarbonyl-2-(N-allyloxycarbonyl-N-sulfamoylamino )methyl-4 -mercaptopyrrolidine (3.77 g: 9.94 mmole) are reacted to give (lR,5S,6S)-2- 5S)-l-allyloxycarbonyl-5-(N-allyloxycarbonyl -Nsulfamoylamino )methylpyrrolidin-3-yl] thio-6- [(iR hydroxye-thyl] -1-methyl- 1-carba-2-penem-3-carboxylic acid allyl ester (3.65 Yield: IR V2 (CHCl 3 cmnf 1 1777, 1718, 1686, 1395.

NMR 5 (CDCl 3 ppm: 1.27(3H, d, J=7.2Hz), 1.37(3H-, d, *0.64J=6.2H-z), 2.5 to 2.7('LH, in), 3.1 to 3.3(3H, in), 3.6 to 9 3.8(2H, in), 4.0 to 4.3(4h, in), 4.4 to 4.9 (6H, in), 0 49..

5.2 to 5.5(6H, mn), 5.7 to 6.1(5H, in).

Step 4 Deprotection Under similar reaction condition to that in :Step 4 in Example 13, (lR,5S,6S)-2-[(3S,5S)-l-allyloxy- )methyl1pyrrolidin-3-yl]thio-6-[(IR )-l-hydroxyethyl] -1-methyll-carba-2-penem-3-carboxylic acid allyl ester (369 mng: 0. 586 minole) is deprotected. with triphenylphosphine (83 mg: 0.32 inmole), tri-n-butyltin hydride (0.64 ml: 9 2.3 minole), and palladium tetrakis(triphenylphosphine) (122mg: 0.11 minole) to give (1R, 5S,6S)-6-t(1R)-lhydroxyethyl]-2-[(3S,5S)--5-sulfamidomethylpy.rrolidin- 3-yllthio-l-methyl-1-carba-2-penem-3-carboxylic- acid (206 mg) Yield: 84%. (HPLC purity :8556) 120 NMR 6 (D20) ppm: 1.22(d, J=7.2Hz, 3H), 1.27(d, J=6.3Hz, 3H), 1.64 to 1.82(m, 1H), 2.62 to 2.80(m, 1H), 3.26 to 3.59(m, 5H), 3.63 to 3.76(m, 1H), 3.84 to 4.10(m, 2H), 4.16 to 4.29(m, 2H).

IR (KBr) cm- 1 3400, 1750.

Example A solution of (1R,5S,6S)-6-[(lR)-1hydroxyethyl]-2-[ (3S,5S)-5-sulfamidomethylphrrolidin- 3-yl]thio-l-methyl-l-carba-2-penem-3-carboxylic acid g) in aqueous sodium hydrogen carbonate (5 ml) at pH 7.0 is filled in a vial (10 ml) and lyophilized.

The lyophilizate is dissolved in water for infection ml) before use and injected thrice a day intrave- 15 nously to a patient suffering from urinary tract infection caused by a sensitive strain of Staphylococcus aureus to cure the disease.

A a Example 16 A solution of (1R,5S,6S)-6-[(lR)-1-hydroxyethyl]-2-[(3S, idin-3-yl]thio-l-methyl-l-carba-2-penem-3-carboxylic acid (0.5 g) in aqueous sodium hydrogen carbonate s.

(5 ml) at pH 7.0 is filled in a vial (10 ml) and lyop- 25 hilized. The lyophilizate is dissolved in water for infection (5 ml) before use and injected thrice a day .i intravenously to a patient suffering from pneumonia :caused by a sensitive strain of Klebsiella pneumoniae to treat the disease.

Example 17 A solution of (1R,5S,6S)-6-[(1R)-l-hydroxyethyl]-2-[(3S,5S)-5-(2-hydroxyethylsulfamoyl)amino- 121 methylpyrrolidin-3-yl]thio-l-methyl-l-carba-2-penem-3carboxylic acid (2.0 g) in aqueous sodium hydrogen carbonate (10 ml) at pH 7.0 is filled in vial (100 ml) and lyophilized. The lyophilizate is dissolved in water for injection (50 ml) before use and administered by infusion four times a day intravenously to a patient severely suffering from the respiratory tract infection caused by a sensitive strain of Enterobacter cloacae to cure the disease.

Various other modifications will be apparent .to and can be readily made by those skilled in the art without departing from the scope and spirit of this invention. Accordingly, it is not intended that the 15 scope of the claims appended hereto be limited to the description as set forth herein, but rather that the claims be broadly construed.

The entire disclosure in the complete specification of our Australian Patent Application No.

21090/92 is by this cross-rrterence incorporated into 0**o the present specification.

C

Claims (4)

1. A pyrrolidine derivative represented by formula II: R4 I R 2 ylS NS 2 N< R3 R\-Y2 II (II) wherein R2, R 3 and R 4 are hydrogen, lower alkyl which can be substituted, or an amino protecting group independently, or R 2 and R 3 together with a nitrogen atrrm to which R 2 and R 3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or R 3 and R 4 together with two nitrogen atoms and one sulfur atom in the f: al eh group form a saturated or unsaturated cyclic group; each cyclic group can further include at least one atom selected from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; yl is hydrogen or a mercapto protecting group; and Y 2 is hydrogen or an amino protecting group.

2. A pyrrolidine derivative according to Claim 1, wherein R 4 is hydrogen.

A method for producing a pyrrolidine derivative represented by Formula II: f. 4 R 2 II R wherein R 2 R 3 and R 4 are hydrogen, lower alkyl which can be substituted, or an amino protecting group independently, or R 2 and R 3 together with a nitrogen atom to which R 2 and R 3 are bonded form a saturated or unsaturated cyclic group, or R and R r R 3 and R 4 together with two nitrogen atoms and one sulfur atom in the f \group form a saturated or unsaturated cyclic stafnkoee JRTYPES2109.9ZIV 17.8 I law 123 grrup; each cyclic group can further include at least one ecom selected from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; Y 1 is hydrogen or a mercapto protecting group; and Y 2 is hydrogen or an amino protecting group; the method comprising the steps of: converting a hydroxy group at the 4-position of a

4-hydroxypyrrolidine-2-carboxylic acid, derivative into a mercapto group; converting a carboxy group at the 2-position into a hydroxymethyl group; converting a hydroxy group in the hydroxymethyl group into an amino group; and o* converting the amino group into a sulfamido group. 4. A method according to Claim 3, wherein R 4 is hydrogen. DATED this 18th day of August 1994 SHIONOGI SEIYAKU KABUSHIKI KAISHA By Their Patent Attorneys: 50 GRIFFITH HACK CO Fellows Institute of Patent e Attorneys of Australia stalvnalkeoqJRETYPES21090.92.IV 17.8 I 124 ABSTRACT A pyrrolidine derivative represented by formula II: R4 2 I R 2 Y1S NSO2N R 3 wherein R 2 R 3 and R 4 are hydrogen, lower alkyl which can be substituted, or an amino protecting group independently, or R 2 and R 3 together with a nitrogen atom to which R 2 and R 3 are bonded form a saturated or unsaturated cyclic group, or R 2 and R 4 or R 3 and R 4 together with two nitrogen atoms and one sulfur atom in the -aroup form a saturated or unsaturated cyclic group; each cyclic group can further include at least one atom se: acted from the group consisting of oxygen, sulfur and nitrogen, and each cyclic group can be substituted; Y1 is hydrogen or a mercapto protecting group; and Y2 is hydrogen or an amino protecting group, useful as intermediates of pyrrolidine derivatives. goes 9 9 stafuna/ke9JWEuYPES12109.92.DIV 18.8 le I