NO841442L - NAFTYLGYLYCYL AND TETRAHYDRONAPHYLYLYLYCYLPHALOSPORINE DERIVATIVES AND PROCEDURES FOR THEIR PREPARATION - Google Patents

Description

The present invention relates to a new group of cephalosporins. which are orally effective and which have advantageous pharmacokinetic properties.

Cephalosporin antibiotics have been very thoroughly investigated,

and several compounds of this group of antibiotics are routinely used to combat bacterial diseases caused by a wide spectrum of both gram-positive and gram-negative microorganisms. Most such compounds are not orally effective, but are administered instead intramuscularly or intravenously, necessitating the assistance of medically trained personnel. Because the compound is effective against a wide spectrum of microorganisms, the compound is not usually used to achieve a specific effect.

Consequently, there is a need for cephalosporin antibiotics which

are orally effective and which have a certain specific effect against one or more groups of microorganisms. The present invention provides a group of compounds which fulfill the above-mentioned requirements.

The present invention thus provides a 2-naphthyl-glycylamido-cephalosporin of formula I:

where

R 1 is in which R 7 and R 8 independently of each other are hydrogen, halogen, hydroxy, C 1 -C 4 -alkyl, C 1 -C 4 -alkyl, nitro, amino, c±~ c^~ alkanoylamino, C^- C 1 -alkylsulfonylamino, or when R 1 and R 2 are joined together, they can form a methylenedioxy group;

where A and B are both hydrogen, or together may be a complete double bond;

R 2 is hydrogen, an amino protecting group, hydroxy or

3 2 3

methoxy, and R is hydrogen, or where R and R together can form:

where M and L can independently be C 1 -C 4 -alkyl group;

4

R is hydrogen, methoxy or methylthio;

R<5> is hydrogen, methoxy, methyl, halogen or methoxymethyl; R 1 is hydrogen, or a carboxy protecting group;

provided that R 2 is only hydroxy or methoxy when A and B form a complete double bond, and that A and B are both hydrogen when R is different from hydrogen; or pharmaceutically acceptable salts of such compounds.

Such compounds are orally active antibiotics, or can be used as intermediates for the production of such antibiotics.

Preferred compounds provided by the present invention include those of formula I wherein R 1 is

and R 7 and R 8 are as defined previously. Within this group, the preferred compounds will be those where R 2 is hydrogen, an amino protecting group, hydroxy or methoxy, and Rc is hydrogen or a carboxy protecting group. Another preferred group of compounds are those where R 1 is

and R 7 ' and R 8 are as defined previously. Particularly preferred compounds within this group include those wherein A,

B, R<2>, R<3>, R4 and R6 are all hydrogen.

A particularly preferred group of compounds within the present invention is defined by means of the following formula:

rc n

where R , R and R are as previously defined. The most preferred compounds are those where R 7 is hydrogen, halogen, 5 6 hydroxy or methoxy, R is methyl or chlorine, and R is hydrogen, and pharmaceutically acceptable salts of such compounds, e.g. sodium or potassium salts.

The present invention also provides pharmaceutical preparations which include a naphthylglycylamido-cephalosporin derivative of formula I, or a pharmaceutically acceptable salt of such a compound, together with pharmaceutical carriers, diluents or the like. A preferred preparation is suitable for oral administration.

The present invention also provides a method for combating bacterial infections in both animals and humans, which includes administering to the patient or the animal in need of treatment, an effective amount of an antibacterial compound of formula I or a pharmaceutically acceptable salt of such a compound. In a preferred method of treatment, naphthylglycylamido-cephalosporin derivatives will be administered orally for the treatment of diseases caused by gram-positive bacteria.

Furthermore, the invention provides a method for producing a compound of formula I, which includes:

A) Acylates a compound of formula II

with an acylating agent of formula III or an activated derivative of such a compound, where A, B, R"*",R<2>, R4, R^ and R^ are -as defined above, and where one subsequently removes - any amino or carboxyl protecting group; B) deblocks a protected acid of formula I wherein R 1 is a carboxy protecting group, thereby providing a compound of formula I wherein R is hydrogen; C) removes an amino-protecting group R 2 from a compound of formula I, whereby a compound of formula I is obtained where R 2 is hydrogen; D) and when it is desired to produce a compound where

2 3

R and R together form a group with formula:

reacts a compound of formula I where R 2 and R 3 are both hydrogen, with a ketone of the formula:

where M and L are as defined above:, or

E) reduces a compound of formula I where A and B together form a double bond and R 2 is hydroxy or methoxy, thereby producing a compound of formula I where A, B and R 2 are hydrogen; and

F) if desired, forms an acid addition salt of

a compound of formula I.

G) or if desired, converts a salt of a compound of formula I into a free amine or a free acid.

In the above formulas, R 1 represents a 2-naphthyl or 2-tetrahydronaphthyl group. Naphthyl and the tetrahydronaphthyl group can be unsubstituted, e.g. when R 10 and R 8 are both hydrogen, monosubstituted in 1, 3, 4,. The 5, 6, 7 or 8 positions,.

e.g. when R 8 is hydrogen and R 7 is different from hydrogen; or disubstituted when R 7 and R 8 are both different from hydrogen. The groups with which the naphthyl and tetrahydronaphthyl rings may be substituted include hydroxy, C 1 -C 6 alkyl, C 1 -C 4 -alkyl, halogen, nitro, amino, C 1 -C 4 -alkanoylamino, and C 1 -C 4 -C 4 -alkylsulfonylamino.

The term "C-^-C^-alkyl" refers to straight and branched lower alkyl groups such as methyl, ethyl, isopropyl, n-propyl, isobutyl and tertiary butyl. Similarly, "C 1 -C 4 alkoxy" denotes lower alkoxy groups attached to the naphthyl ring through an oxygen vacancy. Typical C 1 -C 4 alkoxy groups include methoxy, ethoxy, n-propoxy, n-butoxy and isobutoxy. The term "halogen" means fluorine, chlorine, bromine and iodine. A preferred halogen group is chlorine. By C-^-C^-alkanoylamino is meant an acyl residue of lower alkanoic acid bound to the naphthyl or tetrahydronaphthyl ring through a nitrogen atom. Such groups include formylamino, acetylamino and butyrylamino.

By "C-^-C^-alkylsulfonylamino" is meant a group such as methylsulfonylamino, ethylsulfonylamino and n-butylsulfonyl-

amino.

R 2 represents a substituent on the glycyl nitrogen atom, and may include hydrogen and an amino protecting group. By the term "amino protecting group" is meant any of the well-known substituents which can be attached to an amino nitrogen atom, and which can be easily removed when this is desired. Such protective groups are often used during the preparation of a compound according to the present invention, and serve to reduce the probability of unwanted side reactions that may occur as a result of a free amino group. Although compounds where R 2 is a protecting group will have a biological activity, it is believed that the most biologically desirable compounds will be

and 2

those where R is hydrogen. The compounds where R is an amino-protecting group are thus primarily used as intermediates for a synthesis of the more preferred free amino compounds.

The exact types of amino protecting groups used are

not critical, and one can use any of the well-known protecting groups in this respect. Typical amino protecting groups are

. described by J.W. Barton in "Protective Groups in Organic Chemistry," J.F. McOmie, Ed., Plenum Press, New York, N.Y., 1973, Chapter 2 and by Green in "Protective Groups in Organic Synthesis", John Wiley&Sons, New York, N.Y., 1981 Chapter 7. Both of these references are incorporated herein by virtue of their description of amino protecting groups.

The most common amino-protecting groups that can be used include C₁-C₂-alkanoyl and halogen C₂-C₂-Q-alkanoyl groups such as formyl, acetyl, chloroacetyl, dichloroacetyl, propionyl, hexanoyl, 3,3-diethylhexanoyl, or y- chlorobutyryl, C₁-C₁-alkoxycarbo-

nyl and C 2 -C 4 -alkenyloxycarbonyl groups such as methoxycarbonyl, tertiary butoxycarbonyl and allyloxycarbonyl, C 1 -C 4 ,--aryloxycarbonyl and arylalkenyloxycarbonyl such as benzyloxycarbonyl 4-nitrobenzyloxycarbonyl and cinnamoyloxycarbonyl, halogen-C 4 - C 10 -Alkoxycarbonyl such as 2,2,2-trichloroethoxycarbonyl; and C 5 -C 1 - arylalkyl and alkenyl groups such as benzyl, phenethyl, trityl or allyl.

Other commonly used amino protecting groups include en-amines prepared by reacting the free amino compound with a 6-ketoester, such as methyl or ethyl acetoacetate.

In addition to representing hydrogen or an amino-protecting group, R 2 in the above-mentioned formulas can also together with R 3 form a ring system, so that compounds with the following formula are produced:

14 5 where R , R , R , Rb, M and L are as defined earlier. Examples of such compounds are acetonides, i.e. those where M and L are both methyl. Such compounds can be produced by rea-2 3

generates a glycylamidocephalosporin where R 1 and R 2 are both hydrogen, with a ketone, e.g. acetone. Such cyclic compounds are particularly advantageous as long-acting antibacterial agents.

R in the above formula is hydrogen, an acid addition salt cation,

such as ammonium or an alkali metal cation such as lithium, sodium or potassium, or a carboxy protecting group. With be-

the term "carboxy-protecting group" means any well-known group used to block or protect the carboxylic acid function in a cephalosporin compound during chemical reactions involving other functional positions in the molecule

cooled, and where said group can be easily removed when this is desired using known techniques, e.g. hydrolysis or hydrogenolysis.

Typical carboxy protecting groups used in the present invention include those described by E.

Haslam in "Protective Groups in Organic Chemistry", as above-

for, ch. 5 and by Green in "Protective Groups in Organic Synthesis", reference as above, ch. 5, which is incorporated herein by reference. Examples of commonly used carboxy protecting groups are C-L-C10 alkyl groups such as methyl, tert-butyl, decyl; halo-<C>1<-C>1Q-alkyl, such as 2,2,2-trichloroethyl, and 2-iodoethyl; c5~c15~arylalkyl, such as benzyl, 4-methoxybenzyl, 4-nitrobenzyl, di-

phenylmethyl, C 1 -C 10 alkanoyloxymethyl such as acetoxymethyl or propionoxymethyl; and other groups such as phenacyl, 4-halophenacyl, allyl, dimethylallyl, tri (C 1 -C 2 -alkyl)silyl, such as trimethylsilyl and similar groups.

The naphthylglycyl and tetrahydronaphthylglycyl cephalosporins provided by the present invention can be prepared by one of several methods, one of which involves coupling a 7-aminocephalosporin core of formula II

4 5 6 where R , R .and R are as defined previously, to a naphthyl-glycine compound of formula III or an activated derivative of such a compound:

where A, B, R1, R2, R4, R5 and R6 are as defined above. Examples of cephalosporin cores that can be used in the synthesis of the compound according to the present invention may include

4 5 6

those where R , R and R have the following meaning:

The 7-aminocephalosporin carriers that can be used in the synthesis of compounds according to the present invention are well known and easily accessible by means of known methods. For example 3-halo-cephalosporin cores are available by the methods described in US Patent No. 3,925,372. The 3-methyl-cephalosporin core can be prepared by a ring expansion of the penicillin sulfoxide and with a subsequent side-chain cleavage, or by a hydrogenation of 3-acetoxymethyl derivatives.

The term "activated derivative" means a derivative which makes the carboxyl function in the acetylating agent of formula III reactive with respect to coupling with a primary amino group in order to thereby form the amide bond which binds the acyl side chain to the nucleus. Suitable activated derivatives, methods for their preparation and for their use as acylating agents for a primary amine are well known. Preferred activated derivatives are the following: (a) an acid halide such as the chloride or bromide, or (b) the alkanoyloxy derivative such as formyloxy or acetoxy mixed anhydrides (eg when Y in the following list is HCHO or OCOCH 3 ). Other methods of activating the carboxyl function may involve a reaction between the carboxylic acid and a carbodiimide (e.g. N,N<1->dicyclohexylcarbodiimide, or N,N<1->diisopropylcarbodiimide), thereby obtaining a reactive intermediate which is reacted in situ with the 7-amino group. This reaction is described in more detail below.

In a similar way, the naphthylglycyl and tetrahydronaphthylglycyl reactants as defined by formula III are well known and can be prepared using known methods. Typical naphthylglycyl and tetrahydronaphthylglycyl derivatives that can be used for the preparation of compounds according to the present invention with the above-mentioned formula are those where R<2> is 7, and R 8 can have the following meaning: The coupling of a naphthylglycine or tetrahydronaphthylglycine derivative with a 7-aminocephalosporin nucleus can be carried out using known acylation techniques. For example can a naphthyl-glycylacylating agent, then e.g. the acid chloride or bromide or an alkanoyloxy derivative such as formyloxy or acetoxy anhydrides is reacted with a cephalosporin nucleus, using standard acylation conditions. During such acylation reactions, it is usually preferred that R 2 is an amino protecting group and that R 1 is a carboxy protecting group. Such protective groups will prevent unwanted side reactions as little as possible and they will also increase the solubility of the respective reactants.

The acylation reaction itself is usually carried out by mixing approximately equimolar amounts of a naphthylglycyl or tetrahydro-naphthylglycyl acylating agent of formula III (e.g. an acid halide or a mixed acid halide) with 7-aminocephalosporin iron. The acylation reaction is usually carried out in a mutual solvent, such as benzene, chloroform, dichloromethane, toluene, N,N-dimethylformamide, or acetonitrile,

and will usually be complete after a period of from 1

to 12 hours when carried out at temperatures from -2 0 to + 6 0°. If desired, an equimolar amount can be added

of bases such as pyridine, trimethylamine, aniline or sodium carbonate as an acid adsorbent. The product can be isolated from the reaction mixture by removing the solvent, e.g. by evaporation under reduced pressure, and further purification can, if desired, be carried out by means of routine techniques such as chromatography, crystallization, solvent extraction and other corresponding methods.

An alternative and preferred method for coupling a naphthylglycyl or tetrahydronaphthylglycyl derivative to a 7-aminocephalosporin core in order to thereby obtain compounds according to the present invention, uses a coupling reagent of the type that is routinely used in the synthesis of peptides. Typical coupling reagents that can be used include the carbodiimide such as N,N'-diethylcarbodiimide, N,N-diisopropylcarbodiimide and N,N-dicyclohexylcarbodiimide (DCC); carbonyl coupling reagents such as carbonyldiimidazole; isoxazolinium salts so

such as N-ethyl-5<1->phenylisoxazolinium-3'-sulfonate, and quinoline compounds such as N-ethoxy-carbonyl-2-ethoxy-1,2-dihydro-quinoline (EEDQ).

The coupling of a 7-aminocephalosporin core with a naphthylglycyl or tetrahydronaphthylglycyl derivative using a peptide coupling reagent is usually carried out by mixing approximately equimolar amounts of a 7-aminocef-3-em-4-carboxylic acid derivative, a naphthylglycyl derivative and a peptide coupling reagent, whereby a reaction is obtained according to the following form:

where R , R , R , R and R are as defined above. During such coupling reactions, R 2 is preferably an amino protecting group and R c is hydrogen or a carboxy protecting group. Any protective group present can be removed using the standard method, whereby an active antibiotic according to the present invention is obtained.

The coupling reaction is normally carried out in a mutual solvent such as dichloromethane, acetone, water, acetonitrile, N,N-dimethylformamide or chloroform, and will routinely be fully

continuously over a period of time from 10 to 90 minutes at temperatures from -20 to + 6 0°. Longer reaction periods are not harmful and can be used if this is desired. the product,

a naphthylglycyl or tetrahydronaphthylglycyl cephalosporin, can be isolated by removing the solvent, e.g. by

evaporation under reduced pressure. The product can be further purified using known techniques, e.g. by acid-base extraction, chromatography, salt formation, etc.

Another method for producing compounds according to the present invention uses a naphthyl oxime with the formula

or an activated derivative of such a compound, where R is as defined above, and R 2 is hydroxy or methoxy. When R 2 is hydroxy, this group will usually be protected with a group such as trimethylsilyl or a similar hydroxy protecting group. Such naphthyloxime derivatives can be linked to a cephalosporin nucleus using the methods described above, whereby a compound of formula

where R"*", R<4>, R^ and R^ are as previously defined. These compounds can be used as intermediate products because they can be easily reduced using known methods, whereby preferred naphthylglycylamido compounds according to the present invention are obtained. In addition to this, oximes with the above formula where R is hydrogen, or a salt of such a compound, could be used as antibiotics.

Compounds which have a nitro group on the naphthylglycyl or tetrahydronaphthylglycyl side chain can be modified so that other compounds according to the invention are produced. For example the nitro substituent can be reduced by means of known reduction techniques or by means of hydrogenation, whereby the corresponding amino-substituted naphthylglycylcephalosporin derivative is produced, and if desired, the amino group can be acylated by a reaction with a C^-C^-alkanoyl halide or anhydride or a C 1 -C 2 -alkylsulfonyl halide, which will give the corresponding alkanoylamino or alkylsulfonylaminonaphthyl-glycylamido and tetrahydronaphthylglycyl-amidocephalosporins according to the present invention.

In a similar way, connections can be made according to the present invention

2 3

where R and R together form the group

is produced by reacting a ketone with the formula

with a compound according to the present invention where R 2 and R 3 are both hydrogen, usually in the presence of an acid such as methanesulfonic acid or the like. They produced cyclic compounds, e.g. the preferred acetonides where M and L

both are methyl, are particularly advantageous as oral antibiotics because they are effective for long periods of time.

Other compounds according to the present invention which can be expected to be particularly long-acting antibiotics are those where R<2> is an alkanoylamino protecting group such as formyl or acetyl. Such compounds can conveniently be prepared by simply reacting a naphthylglycylamido-cephalosporin where R 2 is hydrogen with a C-^-C-^Q-alkanoylacylating agent, e.g. formyl chloride or acetic anhydride. Such N-acylated products in themselves will not only act as antibiotics, but will also act as precursor compounds in that they will be hydrolysed in the animal or patient to be treated, to the corresponding naphthylglycyl or tetrahydronaphthylglycyl derivatives.

As the naphthylglycyl and tetrahydronaphthylglycyl side chain in the cephalosporin according to the present invention contains an asymmetric carbon atom, e.g. when A is hydrogen, compounds according to the present invention will exist in the form of optical isomers, namely the D and L isomers. Compounds according to the present invention can be used as a D,L mixture for the treatment of bacterial infections, or if desired, the optical isomers can be separated and used individually. Although both isomers are effective antibacterial agents,

then it appeared that one isomer is more effective than the other, and this was designated the D-isomer and is consequently a preferred embodiment of the invention.

In the case of separation or resolution of the optical isomers can be carried out using known methods which are applied to

a cephalosporin product according to the present invention or on the naphthylglycine or tetrahydronaphthylglycine side chain used as starting compound. A separation of optical isomers can usually be carried out by means of high-pressure chromatography, enzymatic resolution or chemical crystallization or racemisation. A particularly preferred method for producing D-naphthylglycine involves reacting the D,L mixture with benzaldehyde and optically active tartaric acid using the method described in US Patent No. 3,976,680.

As mentioned above, preferred compounds according to the present invention are those where R 2 in the above formula is hydrogen. Such compounds which are primary amides are basic in nature and readily form pharmaceutically acceptable salts on reaction with an acid. The salts which are pharmaceutically acceptable are the preferred salt forms for the treatment of bacterial infections. The term "pharmaceutically acceptable" salts is understood to mean salts that can be used in chemotherapy when treating warm-blooded animals or humans. Typical acids used for the preparation of salts include inorganic acids, such as hydrogen chloride, hydrogen bromide, sulfuric acid or phosphoric acid; as well as organic acids such as acetic acid, trifluoroacetic acid, succinic acid, methanesulfonic acid, oxalic acid or para-toluenesulfonic acid. The connections according to

present invention where both R 2 and R 6 are hydrogen,

readily forms an internal acid addition salt, namely a zwitterion.

Compounds according to the present invention will usually exist as crystalline solids and can be crystallized from known solvents, such as ethanol, water, N,N-dimethylformamide, etc. Furthermore, the compounds where R is hydrogen will be 4-carboxylic acid. Such compounds are acidic in nature and readily form salts with organic and inorganic bases. By the term "pharmaceutically acceptable salts" as used herein, these also include base addition salts.

Compounds according to the present invention will often crystallize as a solvate or hydrate and can be used in this form. As an example, the invention provides a crystalline compound which is 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate. The crystals are large, dense and stable and can be easily ground and sieved for addition to pharmaceutical preparations, especially for solid dosage forms such as filled capsules and the like. The tetrahydrate according to the present invention is prepared by isolating 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid, a preferred product according to the present invention, from an aqueous medium.

This crystalline compound has the following unique X-ray powder diffraction pattern when measured with a 114.6 mm Debye-Scherrer camera containing a nickel-filtered copper radiation of 1.5405Å:

In particular, this compound can be prepared by reacting an acid addition salt of 7-(D-2-naphthyl-glycylamido)-3-methyl-3-cephem-4-carboxylic acid with a base such as sodium hydroxide or triethylamine, so that it is formed corresponding zwitterion which is then crystallized from water.

For example, a salt such as the trifluoroacetic acid salt

or the hydrochloric acid salt is dissolved in water or in a mixture of water and an organic solvent such as acetone or acetonitrile. A base such as aqueous ammonium hydroxide is added to adjust the pH between 3 and 5. The precipitate that forms is the tetrahydrate according to the present invention and this can be easily recrystallized from water.

The above-mentioned compounds according to the present invention can alternatively be prepared by isolating the product of the acylation of 7-amino-3-methyl-3-cephem-4-carboxylic acid (7-ADCA) from a solvent containing water. For example, 7-ADCA, typically a silylated derivative, can be acylated with an N-protected D-2-naphthylglycinic acid halide or a mixed anhydride as described earlier. The acylation can usually be carried out in an organic solvent such as acetonitrile. Once the acylation is complete, protecting groups can be removed by known techniques, and the solution of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid can then be diluted with water so that it contains from 10 to 50% by volume of water, after which the pH of the solution is adjusted to a pH between 3 and 5. The crystalline product produced is 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate.

This crystal form is very stable for long periods of time, but

is nevertheless very well absorbed after oral administration. This is somewhat surprising as the compound is only minimally soluble in water. For example the compound forms a saturated solution in water at 3 to 7°C as indicated in the following table:

Another example of a hydrated form of a compound according to the present invention is 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

The crystals of this compound are large, dense and stable, and can be easily ground and sieved for addition to pharmaceutical preparations, and especially solid dosage forms such as filled capsules and the like. The hydrochloride monohydrate is prepared by crystallizing 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid from an aqueous hydrochloric acid solution.

The above-mentioned crystalline hydrochloride monohydrate has the following unique X-ray powder diffraction pattern when measured with a 114.6 mm Debye-Scherrer camera with a nickel-filtered copper radiation of 1.5405Å:

The hydrochloride monohydrate provided by means of the present invention can be prepared by reacting 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid, usually as a solvate or as the crystalline tetrahydrate described earlier, with hydrochloric acid in water or a solution of water and an organic solvent such as acetone or acetonitrile. Sufficient hydrochloric acid must be used so that the pH in solution remains below 2, usually between 0.1 and 0.8. This reaction will normally be complete within one to three hours when carried out at a temperature between 0 and 60°. The precipitate that forms is the hydrochloride monohydrate and can be easily isolated in a known manner.

Alternatively, the compound can be prepared by isolating the product after the acylation of 7-amino-3-methyl-3-cephem-4-carboxylic acid (7-ADCA) from a solvent containing anhydrous hydrochloric acid. For example 7-ADCA, usually as a silylated derivative, can be acylated with an N-protected D-2-naphthylglycine halide or mixed anhydride. The acylation is usually carried out in an organic solvent, such as acetonitrile. As soon as the acylation is complete, the protecting groups can be removed in the usual way, and the solution of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid can be diluted with water so that it contains from 10 to 50% water by volume, after which the pH of the solution can be adjusted to between 0.1 and 0.8 by adding hydrochloric acid. The. crystalline product formed is 7-(D-2-naphthyl-glycylamido)-3-methyl-cephem-4-carboxylic acid hydrochloride monohydrate.

Examples of typical groups of naphthylglycyl and tetrahydro-naphthylglycyl cephalosporins as well as specific compounds provided by the present invention include those set forth below:

A. Preferred compounds of the formula:

1. R is hydrogen or a salt cation, 7 8

a) R and R are both hydrogen,

5

lal.. R is methyl,

5

let2. R is chlorine,

la3.. R<5> is bromine,

5

let4. R is fluorine,

let5. R<5> is iodine,

5

let6. R is hydrogen,

let7. R is methoxy,

let8. R5 is methoxymethyl,

7 8

b. R is hydrogen and R is 6-methoxy, lbl. R<5> is methyl,

lb2. R<5> is chlorine,

lb3. R^ is motooxy,

lb4. R<5> is methoxymethyl,

7 8 c. R is hydrogen and R is 7-fluorine,

lcl. R 1 is methyl,

lc2. R<5> is chlorine,

lc3. R^ is fluorine,

5

lc4. R is methoxy,

5

lc5. R is methoxymethyl.

7 8

R is hydrogen and R is 6-hydroxy, ldl. R<5> is methyl,

ld2. R<5> is chlorine,

ld3. R 1 is methoxy,

ld4. R^ is bromine,

7 8

e. R is 4-methyl and R is 6-chloro,

sorry. R 1 is methyl,

le2. R5 is methoxy;

laugh3. R 5 is chlorine, or a pharmaceutically acceptable salt

of such compounds.

2. Those with the following formula:

where R"*" is

and R g is hydrogen or a salt cation.

7 8

a. R and R are both hydrogen, ■

2nd floor RJ is methyl,

2a2. R<5> is chlorine,

2a3. R 1 is methoxy,

5

2a4. R is hydrogen.

7 8

b. R is hydrogen and R is 7-hydroxy,

2 pages R<5> is methyl,

2b2. R<5> is fluorine,

2b3. R<5> is iodine,

2b4. R<5> is methoxy,

2b5. R<5> is hydrogen

7 8

c. R is 1-methoxy and R is 8-tert-butyl,

2 cl. R is methyl,

5

2c2. R is methoxy,

2c3. R<5> is chlorine,

2c4. R^ is methoxymethyl,

7 8

d. R is 3-chloro and R is 7-isopropoxy,

5

2 dl. R is hydrogen,

5

2d2. R is methoxymethyl,

5

2d3. R is methoxy,

2d4. R<5> is methyl,

2d5. R<5> is chlorine,

or a pharmaceutically acceptable salt of such compounds.

B. Those with the following formula

å5

1. R is hydrogen, R is methyl,

a. R7 and R8 are both hydrogen, R<2> is tert.-butoxycarbonyl,

lol. R is p-nitrobenzyl,

let2. R<6> is 2,2,2-trichloroethyl,

leave3. R is trimethylsilyl,

let4. R 1 is phenacyl,

b. R and R are both hydrogen, R is tert-butyl,

lb. R<2> is tert,-butoxycarbonyl,

2

lb2. R is acetyl,

lb3. R 2 is p-nitrobenzyloxycarbonyl,

2

lb4. R is chloroacetyl,

c. R7 is hydrogen, R<8> is 7-methoxy, R<2> is 2,2,2-trichloroacetoxycarbonyl,

lcl. R is p-nitrobenzyl,

lc2. R is sodium cation,

lc3. R is methyl,

4 5

2. R is methoxy, R is chlorine,

a. R 7 and R 8 are both hydrogen, R 2 is formyl,

2nd floor R<6> is 2,2,2-trichloroethyl,

2a2. R is hydrogen,

2a3. R is sodium cation,

A7 8

3. R is methylthio, R and R are both hydrogen,

5

a. R is methyl,

3rd floor R<2> is tert-butoxycarbonyl,

3a2. R is para-nitrobenzyl,

b. R is bromine,

3 pages R 2 is tert.-butoxycarbonyl, 3b2- R c is trimethylsilyl,

5

c. R is methoxymethyl,

2

3 cl. R is hydrogen,

2

3c2. R is trimethylsilyl,

3c3. R<6> is allyl,

4 7 8 4. R is methylthio, R is hydrogen and R is 6-methoxy,

a. R^ is methoxy,

2

4 al. R is tert-butoxycarbonyl, 4a2. R 6 is methyl,

b. R<5>is chlorine,

2

4 pages R is hydrogen,

2

4b2. R is phenacyl,

c

4b3. R is tert-butyl,

C. Those with formula:

in which R is

1. R<7>and R 8 are both hydrogen,

a. R^ is methyl,

lol. R g is para-nitrobenzyl,

let2. R<6> is tert-butyl,

g

leave3. R is hydrogen,

let4. R 1 is trimethylsilyl,

b. R^ is fluorine,

lb. R<6> is methyl,

c. R 1 is methoxymethyl.

The following examples illustrate the invention.

Production 1

Preparation of 2-naphthylglycine (also called α-amino-α-(2-naphthyl)acetic acid).

A solution of 15.6 g (0.1 m) of 2-naphthaldehyde in 700 ml of 50% ethanol-water containing 14.7 g (0.3 m) of sodium cyanide and 28.4 g (0.4 m) ammonium carbonate was kept at 50° for 20 hours. The reaction mixture was cooled and concentrated to about 400 ml by evaporation under reduced pressure, after which the solution was acidified to pH 2.0 by adding concentrated hydrochloric acid. The solid precipitate was filtered off, washed with dilute hydrochloric acid and dried, yielding 22.1 g of 4-(2-naphthyl)-2,4-imida-zolidendiones.

A solution of 5.0 g (22 mM) of 4-(2-naphthyl)-2,4-imidazolidinediones in 100 ml of 16% (v/v) aqueous sodium hydroxide was refluxed for 2.5 h. The reaction mixture was then filtered, cooled and washed with ethyl acetate. The aqueous solution was then diluted with 6N hydrochloric acid to pH 5.1 and filtered, yielding 2-naphthyl-glycine. The reaction was repeated several times to produce larger quantities of the product.

A 10.1 g sample of 2-naphthylglycines was purified by at

the sample was dissolved in 125 ml of methanol containing 2.9 ml of acetyl chloride. The reaction mixture was filtered, after which the filtrate was diluted with 5 ml of aniline. The precipitated product was filtered off and dried, which gave 7.0 g of 2-naphthylglycine,

sm.p. 219-221°C.

Manufacturing 2

Dissolution of 2-naphthylglycine.

A mixture of D and L 2-naphthylglycines was treated with optically pure α-aminoethylbenzene in the presence of N,N'-dicyclohexylcarbodiimide, whereby N-(1-phenylethyl)-α-amino-α-(2- naphthyl)acetamide. A separation of the diastereomers by chromatography over silica gel gave, after acid hydrolysis, D-2-naphthylglycine (OR -190° + 3°) and L-2-naphthylglycine (OR = + 190° - 3°).

Manufacturing 3

Preparation of 6-methoxynaphth-2-yl-glycine.

2-Bromo-6-methoxynaphthalene was converted to 2-lithio derivatives by reaction with n-butyllithium. The diethyl oxalate was then reacted with said 2-lithio-6-methoxynaphthalene, which gave ethyl α-keto-6-methoxy-naphth-2-yl acetate. The latter connection was re-

added with hydroxylamine hydrochloride and sodium acetate to give ethyl α-hydroxyimino-6-methoxynaphth-2-ylate. A solution of 17.55 g of the oxime in 600 ml of methanol containing 5.3 g of zinc metal dust and 135 ml of 50% (v/v) aqueous formic acid was stirred at 0° for three hours. After filtering the reaction mixture, solution was removed by evaporation, yielding 10.3 g of ethyl α-amino-α-(6-methoxynaphth-2-yl)acetate.

NMR (CDCl 3 ). 61.20 (t, 3H),6 2.15 (s, 2R\,6 3.89 (s, 3H),

<5 4.15 (m, 1H) , 6 4.72 (s, 1H) , 66.08-6.75 (m, 6H) .

Hydrolysis of the produced ester by reaction with 1N sodium hydroxide gave 6-methoxynaphth-2-ylglycine.

Manufacturing 4

Preparation of N-tert.-butoxycarbonyl-2-naphthylglycine.

To a stirred solution of 10 g (50 mM) of 2-naphthylglycine (from Preparation 1) in 100 ml of 1N sodium hydroxide was added 50 ml of tetrahydrofuran followed by 30 g (140 mM) of di-tert-butyl carbonate. The reaction mixture was stirred at 24°C for 4

hours. The product was isolated by first washing the reaction mixture three times with 50 ml portions of diethyl ether, after which the mixture was acidified to pH 2.0 by adding conc. hydrochloric acid. The aqueous acidic mixture was extra-

quenched several times with ethyl acetate, the extracts were combined, washed with water, dried, after which the solvent was removed by evaporation under reduced pressure to give 12.8 g (85% yield) of N-tert-butoxycarbonyl-2-naphthyl -glycine. NMR (DMSO-dg): δ 2.5 (s, 9H), δ 6.85 (s, 1H), δ 7.28-7.9 (m, 7H).

By using the methods indicated in preparations 1-4, the following compounds were prepared:

N-tert.butoxycarbonyl-(6-methoxy-2-naphthyl)-glycine,

(NMR (CDC13): 6 1.2 and 1.4 (two broad singlets) 9H), 6 5.4 (broad singlets, 1H), 6 6.7 (broad singlets, 1H), 6 7.03-7 .8 (m, 6H).

N-tert.-butoxycarbonyl-(6-hydroxy-2-naphthyl)-glycine,

NMR (CDCl 3 ), δ 1.2-1.4 (broad singlet, 9H), δ 5.3-5.9 (two broad singlets, 1H), δ 6.9-8.5 (m, 7H).

N-tert-butoxycarbonyl-(6-chloro-2-naphthyl)-glycine.

NMR (CDCl 3 ): δ 1.15 (s, 9H), 65.3-5.7 (m, 1H), δ 7.3-8.3 (m, 8H).

Manufacturing 5

α-amino-ct-(2-naphthyl) acetyl chloride hydrochloride id

Hydrogen chloride was bubbled through a cold 0° solution of 5.0 g

(25 mM) 2-naphthylglycine in 150 mL dichloromethane for 20 min. The reaction mixture was thus stirred while 7.6 g (38 mM) phosphorus pentachloride was added in one portion, after which the stirring was continued at 0-10° for 2 hours. The solution was filtered, dried, after which the solvent was removed by evaporation under reduced pressure to give 5.2 g 81% yield of α-amino-α-(2-naphthyl)acetyl chloride hydrochloride IR (mull)

1795 cm"<1>

Analysis calculated for C^H-^Cl^O

Theoretical: Cl, 27.68

Found: Cl, 27.69.

Production 6

Enzymatic resolution of D,L 2-naphthylglycine.

Using the general procedure described in US Patent No. 3,386,888, 19.8 g of D,L-N-chloroacetyl-2-naphthylglycine was reacted with 4 g of N-acyl L-amino acid amidohydro-

dissolve in 1250 ml of 0.1 M potassium hydrogen phosphate pH 7.0 buffer inside--4

containing 5 x 10 M cobalt chloride hexahydrate. The reaction mixture was shaken for 2 hours at 27°C. L-2-naphthylglycines had at this point precipitated and were filtered off. The filtrate was acidified to pH 2 by the addition of 1N hydrochloric acid, after which the mixture was extracted twice with 500 ml portions of ethyl acetate. The extracts were combined, dried, after which the solvent was removed by evaporation under reduced pressure to give 9.065 g of D-N-chloroacetyl-2-naphthylglycine (91.5% yield).

Analysis calculated for C^H-^NO^Cl

Theoretical C, 60.55, H, 4.36, N, 5.04

Found: C, 60.62, H, 4.34, N, 4.76

"212.0° L-2-naphthylglycine which was filtered off was washed with pH 7.0, then with water and then with hexane and then air dried, where-

where 6.82 g (95% yield) of L-2-naphthylglycine were obtained.

Analysis: calculated for C]_2HllN02

Theoretical: C, 71.63, H, 5.55, N, 6.96.

Found: C, 69.85, H, 5.62, N, 6.51.

1~% 5J +195.2°.

Manufacturing 7

8-nitro-2-naphtholic acid.

5-nitro-2-naphtholic acid.

A stirred solution of 400 ml conc. hydrochloric acid, 18 g (0.11 m) of 2-naphtholic acid were added in portions at 60°. The reaction mixture was kept at 70°C for 2 hours, cooled by adding 200 g of ice. The precipitate was filtered off and dried to 18.8 g (77% yield) of a mixture of 5 and 8-nitro-2-naphtholic acid. The mixture was converted to the ethyl ester by reaction with ethanol in the presence of sulfuric acid. 5 g of the mixture of ethyl esters were crystallized from 20 ml of ethyl acetate, whereby 800 mg of ethyl 8-nitro-2-naphthoate m.p. 120° and 1.9 g of ethyl 5-nitro-2-naphthoate.

Manufacturing 8

Ethyl 8-amino-2-naphthyl formate.

A solution of 11.2 g of ethyl 8-nitro-2-naphthoate (prepared

as described in preparation 7) in 100 ml of ethanol was hydrogenated in the presence of 5% palladium on carbon. The reaction mixture was filtered and the solvent removed from the filtrate to give ethyl 8-amino-2-naphthyl formate.

Production 9

Ethyl 8-hydroxy-2-naphthyl formate.

A solution of 3.1 g ( 45 mM)

of sodium nitrite in 25 liters of water. The reaction mixture was stirred for 50 minutes and then a hot solution (90°C) of 90 ml of water in 10 ml of conc. sulfuric acid. The reaction mixture was stirred for 10 minutes at 90°, then cooled and then extracted with di-

chloromethane. The extracts were combined, washed with salt solution, and dried, after which the solvent was removed and after waiting by chromatography, 1>7 g of ethyl 8-hydroxy-2-naphthyl formate, sm. p. 136-137°C.

Using the same general procedure, 14.9 g of ethyl 8-amino-2-naphthyl formate was reacted with tert-butyl nitrite and copper (II) chloride, which gave 11.5 g of ethyl 8-chloro-2-naphthyl formate.

Production 10

Ethyl 8-methoxy-2-naphthyl formate.

A solution of 216 mg of ethyl 8-hydroxy-2-naphthylformate (from Preparation 9) in 15 ml of acetone containing six drops of dimethyl sulfate and 150 ml of potassium carbonate was stirred at 25° for 24 hours. The solvent was removed and the product was dissolved in ethyl acetate, washed with 5N hydrochloric acid and with brine, then dried and concentrated to 200 mg of ethyl 8-methoxy-2-naphthyl formate.

Production 11

ct-Methoxyimino-α-(8-chloro-2-naphthyl)acetic acid.

A suspension of 9.2 g of sodium hydride in 50 ml of N,N-dimethylformamide was added in one portion to a stirred solution of 5.6 g

(24 mM) of ethyl 8-chloro-2-naphthylformate and 4.4 g (36 mM) of methyl-methylthiomethylsulfoxide in 10 ml of N,N-dimethylformamide. The reaction mixture was stirred for 4 hours at 25°C, and then concentrated to dryness. The product was dissolved in 250 ml of ethyl acetate, and the solution was then washed with 5° hydrochloric acid and then with saturated sodium bicarbonate and saline. The solution was dried and the solvent was removed by evaporation to give 4.73

g (63% yield) of 1-oxo-1-(8-chloro-2-naphthyl)-2-methylthio-2-methylsulfinylethane. A solution of 3.12 g (10 mM) of the product in 150 ml of formic acid and 12 ml of acetic anhydride was stirred at 65°

for \ hour. To the reaction mixture was then added 856 mg (4mM) sodium periodate, and stirring was continued for another quarter of an hour. The reaction mixture was then cooled, and concentrated to dryness

hot, after which the product was dissolved in ethyl acetate, washed with sodium bicarbonate and a saturated sodium chloride solution, after which the solution was removed, giving 1.2 g (46% yield) of methylthio α-oxo-α-(8-chloro-2-naphthyl) )acetate.

A solution of 220 mg of a product prepared as described above in 15 ml of methanol and 15 ml of water containing 0.83 ml of 1N sodium hydroxide and 70 mg of methoxyamine hydrochloride was stirred for 16 hours at 25°. The reaction mixture was acidified to pH 2 by adding 1N hydrochloric acid. The acidic solution was extracted with ethyl acetate and the extracts were dried and concentrated to 170 ml of α-methoxyimino-α-(8-chloro-2-naphthyl)acetic acid.

Using the same procedure, the following compounds were prepared: α-methoxyimino-α-(8-nitro-2-naphthyl)acetic acid,

α-methoxyimino-α-(8-amino-2-naphthyl)acetic acid,

α-methoxyimino-α-(8-hydroxy-2-naphthyl)-acetic acid, and α-methoxyimino-α-(8-methoxy-3-naphthyl)acetic acid.

Production 12

Preparation of 7-amine-3-chloro-3-cephem-4-carboxylic acid.

US Patent No. 2,925,372 describes the synthesis of 7-phenylglycyl-amido-3-chloro-3-cephem-4-carboxylic acid, usually generically referred to as cefaclor. The reaction of cefaclor with phosphorus pentachloride, methanol and water under known conditions for cleavage of cephalosporin side chains gives 7-amino-3-chloro-3-cephem-4-carboxylic acid.

In a similar manner, the following cephalosporin cores can be prepared for use during the synthesis of compounds according to the present invention: 7-amino-7-methoxy-3-bromo-3-cephem-4-carboxylic acid, 7-amino-3-cephem-4-carboxylic acid,

7-amino-3-methoxymethyl-3-cephem-4-carboxylic acid, 7-amino-7-methylthio-3-methyl-3-cephem-4-carboxylic acid.

Production 13

7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid N,N-dimethylformamide monohydrate.

To a cold (-20 to -30°) stirred suspension of 51.0 g of D/~N-(1-methoxycarbonyl)-2-propenyl7~2-naphthylglycine sodium salt, prepared by reacting methyl acetoacetate with D-2-naphthylglycine ,

1,500 ml of acetonitrile containing 250 ml of N,N-dimethylformamide,

was added 0.44 ml of methanesulfonic acid, then 0.45 ml of N,N-dimethylbenzylamine and then 12.35 ml of methyl chloroformate. The reaction mixture was stirred for 2 hours at -20 to 30° after the addition. It was then diluted by adding a cold

(0°) solution of 34.24 g of 7-amino-3-methyl-3-cephem-4-carboxylic acid in 230 ml of acetonitrile, containing 59.4 ml of N-methyl-monotrimethylsilyl-trifluoroacetamide. The reaction mixture was stirred for 2 hours at -20 to -30°, and then warmed to 0°. The reaction mixture was acidified by adding 160 ml of 1N hydrochloric acid, after which the mixture was stirred while adding 17.85 g of semi-carbazide hydrochloride in one portion. pH

on the mixture was adjusted to 3.0 and held at this level by the addition of triethylamine. After the mixture was heated to 25°C, it was filtered through hyflo filter aid. The filtrate was diluted by addition of triethylamine to pH

6.2 and cooled to 0° for 45 minutes and then filtered. Filter-

the cake was washed 4 times with 50 ml portions of acetonitrile, and dried at 35°C for 7 hours under reduced pressure, yielding 6.8 g of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem- 4-carboxylic acid N,N-dimethylformamide monohydrate.

Yield 86% Karl Fisher Sample: 5.69%,

NMR (TFA): signals at 6 2.1 (s, 3H), 3.2 (S, 3H), 3.3(ABq, 2H), 3.4(s, 3H), 5.2 (d, 1H), 5.8 (dd, 1H), 5.8(S, 1H), 7.3-8.2(M, 7H), 8.3 (S, 1H).

Example 1

7-(2-Naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid trifluoroacetate salt.

A stirred suspension of 1.0 g of 4.7 mM 7-amino-3-methyl-3-cephem-4-carboxylic acid (7-ADCA) in 25 ml of acetonitrile was added in one portion to 3.7 ml of 14.0 mM ( trimethylsilyl)trifluoroacetamide. The reaction mixture was stirred at room temperature until all solids had dissolved, indicating complete formation of the trimethylsilyl ester of 7-ADCA.

In a separate flask, a solution of 1.35 g (4.5 mM) of N-tert.-butoxycarbonyl-2-naphthylglycine (from preparation 4) was stirred

in 20 ml of acetonitrile containing 1.1 g (4.5 mM) N-ethoxy-carbonyl-2-ethoxy-1,2-dihydroquinoline (EEDQ) at room temperature

trip for a quarter of an hour. This solution was then added in one portion to

added the cold (0°C) acetonitrile solution containing the trimethylsilyl ester of 7-ADCA prepared as described above. The reaction mixture was stirred for 1 hour at 0° and heated

to room temperature. The solvent was removed by evaporation under reduced pressure to give an oil which was dissolved in ethyl acetate, washed twice with 1N hydrochloric acid, then dried,

after which the solvent was removed by evaporation to give 7-(N-tert-butoxycarbonyl-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid as a foam.

The N-protected naphthylglycyl cephalosporin thus prepared

was dissolved in 5 ml of trifluoroacetic acid, after which this acid was removed by evaporation under reduced pressure, and after precipitation from diethyl ester, 5.7 g of 7-(2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid trifluoroacetate salt was obtained .

Example 2

7-(2-Naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate.

A mixture of 5.7 g of the trifluoroacetic acid addition salt from Example 1 in 55 ml of 10% (v/v) water and acetonitrile was heated to approx. 50°, and then filtered to remove undissolved solids. The filtrate was then diluted with 1.8 molar ammonium hydroxide to pH 4.5. The precipitate was filtered off and dried to give 3.15 g (72% yield) of 7-(2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate.

Example 3

7-(D-2-Naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate.

The procedure from Example 1 was repeated, but using 5.0 g of optically active D-N-tert.-butoxycarbonyl-2-naphthylglycine and 5.6 g of 7-ADCA, which gave, after removing the N-protecting group, 6, 8 g (80% yield) D-'7-(2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid trifluoroacetate salt.

The prepared salt was dissolved in 90 ml of acetonitrile and

10 ml of water containing 5 ml of triethylamine. The reaction mixture was stirred at 25° for 20 minutes and then filtered. The filtrate was concentrated to dryness and the product was crystallized from water to give 2.9 g of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate, m.p. 171-180°C (decomp.).

Analysis: calculated for<C>^<H>^<N>^<OgS>

Theoretical: C, 51.16, H, 5.80, N, 9.95, S, 6.93.

Found: C, 52.52, H, 5.47, N, 9.73, S, 6.83.

NMR (DMS0-d6) δ 1.9 (s, 3H), δ 4.8 (s, 1H), δ 4.9 (dd, 1H),

δ 5.6 (dd, 1H), 67.49-7.99 (m, 7H). Example 4 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate. D-2-naphthylglycine sodium salt was protected as an enamine by reaction with methyl acetoacetate. A suspension of 102 g (317.7 mM) of the protected D-2-naphthylglycine sodium salt in 1000 ml of acetonitrile and 500 ml of N,N-dimethylformamide was cooled at -30° and stirred while adding in one portion 0.88 ml of methanesulfonic acid, and then 0.90 ml of N,N-dimethylbenzylamine and 24.7 ml of methyl chloroformate. The reaction mixture was stirred at -30° for two hours, and then diluted by dropwise addition of a solution of 68.5 g (302.8 mM) 7-amino-3-methyl-3-cephem-4-carboxylic acid in 460 ml of acetonitrile containing 118.8 ml of hexamethyldisilazane. The reaction mixture was stirred at -30° for two hours after the addition was complete, after which it was warmed at 0°C_. It was then diluted by adding 120 ml of 1N hydrochloric acid and then added 35.7 g of semicarbazide hydrochloride. Ammonium hydroxide was added to adjust and maintain the pH at 3.0 while the mixture was heated at 22°. The mixture was further diluted by adding 430 ml of water,

and then decolorized by stirring for a quarter of an hour with 10.0 g of charcoal. The reaction mixture was filtered through a hyflo filter aid and the filtrate was heated to 40°. The pH was adjusted to 4.0 by adding IN ammonium hydroxide, whereby crystallization was obtained. The crystallization continued for approx. k hour, after which the pH was raised to 5.2 by adding 1N ammonium hydroxide. The mixture was cooled to 20° and stirred for 1 hour and then filtered. The filter cake was washed twice with 50 ml portions of water, and then air dried to 110.8 g of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate.

Analysis: calculated for C^g<H>^<gN>^<O>^<S>.4H20

theoretical: C, 51.16, H, 5.80, N, 8.97, S, 6.83,

Found: C, 50.31, H, 5.62, N, 8.87, S, 6.89.

Karl Fisher water analysis:

Theoretical: 15.3

Found: 13.73%

NMR (TFA): δ 2.2 (s, 3H), 3.21 (q, 2H), 5.08 (d, 1H), 5.68,

(m, 2H), 7.4-8.3 (m, 7H), 10.5 (s, 12H).

IR (KBr): 1766, 1751, 1696 cm<-1>

UV (CH3 -.OH) : X max 227, e 78,000

A 265, e 12,000

max

Titration (65% N,N-dimethylformamide/water)

pKa 5.5, 7.5, 11.2.

Example 5

7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

A stirred solution of 200 ml of water containing 10 ml of 12N hydrochloric acid was added in one portion to 20 g of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid N,N-dimethylformamide solvate (from production 13). The reaction mixture was adjusted to pH 0.6 by adding 12N hydrochloric acid and then stirred at 25°

for 45 minutes. The precipitated solid was filtered off and washed once with 50 ml of water. The crystalline material was then dried at 35 to 40° for 16 hours, yielding 17.69 g (92%) of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

Analysis: calculated for C2qH22<C>1N20^S

Theoretical C, 53.15, H 4.91, N, 9.30, S, 7.09, O, 17.70,

Cl, 7.85

Found: C, 52.75, H, 4.85, B, 9.85, S, 7.03, 0, 17.07,

Cl, 7.60.

Karl Fisher analysis:

Theoretical: 4.29%

Found: 4.62%.

Chloride test (by filtration): theoretical: 7.85, Found 8.16

and 8.02. IR (KBr) 3040, 2940, 1768, 1707, 1533, 1232 cm"<1>.

UV (EtOH): X 225, e = 100,000.

max

Titration (66% N,N-dimethylformamide-water v/v): 5.5, 7.3

Example 6

7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

A solution of 5 g (10.6 mM) 7-(D-2-naphthylglycamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate (from Example 3)

in 50 ml of acetone was heated to 40° and then acidified by adding 1 ml of 12N hydrochloric acid. The mixture was stirred and diluted by adding 5 ml of water and 6N ammonium hydride to pH 3.3.

The pH was lowered to 0.2 by dropwise addition of 12N hydrochloric acid,

after which the reaction mixture was stirred for 2 hours at 25°.

The precipitated product was filtered off, washed 2 times with

10 mL of water and dried at 25° under vacuum for 2 hours to give 1.929 g (40.5% yield) of crystalline 7-(D-2-naphthylglycylamido)-3-methyl-3-dephen-4-carboxylic acid hydrochloride monohydrate.

Chloride analysis: Theoretical 7.85, - found 7.96%.

Example 7

7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

To a warm (40°C) stirred solution of 1 ml of 12N hydrochloric acid and

25 ml of water and 25 ml of acetone were added in one portion to 5 g of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate (from example 3). Additional 12N hydrochloric acid was added to the reaction mixture to adjust the pH to 0.5, after which the mixture was cooled to 25° and stirred for 2 hours.

The precipitated product was filtered off, washed with three 10 ml portions of water, after which it was dried for 4 hours at 25° under vacuum. The product was found to be 3.596 g (75% yield) of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

Karl Fisher analysis:

Theoretical: 4.2 9%

Found: 5.95%.

Chloride analysis: Theoretical 7.85%, found 8.0 9%.

Example 8

7-(D-2-Naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

A solution of 10 g (21.3 mM) of 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate in 100 ml of water containing 5 ml of 12N hydrochloric acid was stirred at 40° while 12N hydrochloric acid was added dropwise to pH 0.6. The mixture was cooled to 25°C and stirred at said temperature for 1 hour. The crystalline product was filtered off, washed twice with 20 ml portions of water, and then dried for 3 hours at 25° under reduced pressure. The dried product was identified as 8.16 g (84.5% yield) of crystalline 7-(D-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate.

Karl Fisher analysis:

Theoretical: 4.29%

Found: 5.3%.

Chloride analysis: Theoretical 7.85, found 7.62%.

Example 9

p-nitrobenzyl - 7-(N-tert.-butoxycarbonyl-2-naphthylglycylamido)-3-chloro-3-cephem-4-carboxylate.

A solution of 260 ml of EEDQ in 50 ml of acetonitrile containing 301 ml of N-tert.-butoxycarbonyl-2-naphthylglycine was added in one portion to a stirred cold (0°) solution of 682 ml of p-nitrobenzyl 7-amino-3-chloro- 3-cephem-4-carboxylate in 50 ml of acetonitrile. The reaction mixture was stirred for 90 minutes at 0°

and then warmed to room temperature. The solvent was removed by evaporation under reduced pressure to give an oil. This was dissolved in ethyl acetate and washed with dilute hydrochloric acid, pH 7.0/buffer, and finally with sodium chloride solution. After drying, the solvent is removed by evaporation, whereby 5.98 mg 93% yield of p-nitrobenzyl-7-(N-tert-butoxycarbonyl-2-naphthylglycylamido)-3-chloro-2-cephem-4-carboxylate was obtained.

NMR (CDCl 3 ): δ 1.21, (s, 9H), δ 3.1-3.8 (m, 2H). 6 4.9

(two duplicates, 1H), 6 5.28 (s, 2H), 6 5.4-6.2 (m, 3H), 5 7.2-8.21 (m, 12H).

Example 10

7-(2-Naphthylglycylamido)-3-chloro-3-cephem-4-carboxylic acid trifluoroacetate. ■

A solution of 598 mg of p-nitrobenzyl 7-(N-tert-butoxy carbonyl-2-naphthylglycylamido)-3-chloro-3-cephem-4-carboxylate (from Example 9) in 50 ml of tetrahydrofuran containing 10 ml of methanol and 5 ml of ethanol was added in portions 1.20

g 5% palladium on carbon. The reaction mixture was shaken for 1 hour in a Paas hydrogenation flask at an initial hydrogen pressure of approx. 4.0 kg per cm. The reaction mixture was then filtered and the solvent removed from the filtrate by evaporation under reduced pressure to give an oil. This was dissolved in a pH 7.0 buffer, after which the pH was adjusted to 7.75 by adding dilute sodium hydroxide. The aqueous solution was washed with ethyl acetate and diethyl ether,

and then acidified at pH 2.3 by adding 1N hydrochloric acid. The solution was then extracted with methyl acetate, and the extract was dried and the solvent was removed by evaporation to give 360 mg (77% yield) of 7-(N-tert-butoxycarbonyl-2-naphthylglycylamido)-3-chloro-3 -cephem-4-carboxylic acid.

The acid produced was dissolved in 6 ml of trifluoroacetic acid, after which the solution was stirred for 5 minutes at 25°. pH

was then adjusted to 3.9 by adding dilute ammonium hydroxide, after which the solvents were removed by evaporation to give an oil. This was subjected to gradient chromatography over reverse phase silica gel, performing the elution at 0 to 25% acetonitrile, 1% acetic

acid and 99-74% water (v/v). Appropriate fractions were combined and the solvent was removed by lyophilization, yielding 49 mg of L- and 49 mg of D-7-(2-naphthylglycylamido)-3-chloro-3-cephem-4-carboxylic acid.

NMR (TPA-d1): 63.82 (q, 2H), δ 5.49 (d, 1H), δ 5.8-6.1 (m, 2H), δ 7.6-8.35 (m , 7H).

Example 11

7-(2-Naphthylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid trifluoroacetate.

A solution of 1.05 g of D,L-N-tert.-butoxycarbonyl-2-naphthyl-glycine in 15 ml of acetonitrile containing 1.06 g of EEDQ was stirred

at 25° for h hour and then in one portion added a stirred up-

solution of 1.0 g of 7-amino-3-methoxy-3-cephem-4-carboxylic acid in 15 ml of acetonitrile containing 1 ml of bis(trimethylsilyl)trifluoroacetamide. The reaction mixture was stirred for 2 hours at 25° and then concentrated to dryness by evaporation of the solvent under reduced pressure to give an oil. This was dissolved in 10 ml of trifluoroacetic acid and stored in wood

0° for 5 minutes. An evaporation of the solvent and a purification of the product by chromatography over silica

gel gave 89.5 ml of D,L 7-(2-naphthylglycylamido)-3-methoxy-3-cephem-4-carboxylic acid trifluoroacetate.

IR(KBr): 1762.10 cm<-1> (B-lactam),

NMR (TFA-d^) : 6 3.2-4.25 (m, 5H) , 6 5.2-5.9 (m, 3H) , 6 7.5-

8.3 (m, 7H).

Example 12 D-7-(6-Chloronaphth-2-ylglycylamido)-3-methyl-3-cephem-4-carboxylic acid.

2-Chloronaphthalene was acylated with ethyl chlorooxalate to give ethyl-α-keto-α-(6-chloro-naphth-2-yl)acetic acid. A reaction of the latter compound with hydroxylamine, followed by a reduction and hydrolysis, gives 6-chloronaphth-2-ylglycine. This compound was converted to N-tert.-butoxycarbonyl protected derivatives.

To a stirred, cold (0°C) solution of 523 mg (1.5 mM) p-nitrobenzyl-7-amino-3-methyl-3-cephem-4-carboxylate in 300 ml of acetonitrile was added a solution of 500 mg ( 1.5 mM) N-tert-butoxycarbonyl-6-chloronaphth-2-ylglycine in 100 ml acetonitrile containing 36 9 mg EEDQ. The reaction mixture was stirred for 1 hour at 0°, and then warmed to room temperature and stirred in

48 hours. The solvent was removed by evaporation under reduced pressure to give an oil. This was dissolved in 100 ml of ethyl acetate and washed with 1N hydrochloric acid, aqueous sodium bicarbonate, and water, and then dried. Removal of the solvent by evaporation gave 770 mL of a white solid.

(77% yield) of p-nitrobenzyl 7-(N-tert-butoxycarbonyl-6-chloronaphth-2-ylglycylamido)-3-methyl-3-cephem-4-carboxylate.

NMR (CDCl 3 ): δ 1.40 (s, 9H), δ 2.12 (two singlets, 3H),

6 3.40 (q, 2H), 6 4.68 and 4.90 (two duplicates, 1H), <5 5.30

(broad s, 3H), 6 5.6-6.0 (m, 1H), 6 7.2-8.3 (m, 8H).

Removal of the p-nitrobenzyl-carboxy protecting group was

carried out by hydrogenating 770 mg of the compound as described above, with 1.0 g of 5% palladium on carbon in 50 ml of methanol containing 20 ml of ethanol with an initial hydrogen pressure of approx. 4 kg/cm. The reaction was complete after three quarters of an hour, after which the mixture was filtered and the solvent was removed, yielding an oil. This was dissolved in 50 ml of ethyl acetate containing pH 7 buffer, and the aqueous layer was acidified to pH 2.3 with 1N hydrochloric acid. The product was extracted into ethyl acetate, then washed with water, dried and concentrated to dryness to give 120 mg 36% yield

exchange of 7-(N-tert.-butoxycarbonyl-6-chloronaphth.-2-ylglycyl-amido)-3-methyl-3-cephem-4-carboxylic acid.

NMR (CDCl 3 ): δ 1.46 (s, 9H), δ 2.15 (two singlets, 3H),

δ 3.35 (m, 2H), δ 7.2-8.1 (m, 8H).

The product prepared was dissolved in 5 ml of trifluoroacetic acid,

and the solution was stirred at room temperature for 5 minutes.

An evaporation of the solvent and a purification of the pro-

conducted by high pressure liquid chromatography gave D-7-(6-chloronaphth-2-ylglycylamido)-3-methyl-3-cephem-4-carboxylic acid.

Example 13 D-7-(2-naphthylglycylamido)-3-methoxymethyl-3-cephem-4-carboxylic acid.

A solution of 570 mg of diphenylmethyl-7-amino-3-methoxymethyl-3-cephem-4-carboxylate tosylate in 30 ml of ethyl acetate in 30 ml of ethyl acetate containing 10 ml of aqueous sodium bicarbonate was stirred for 5 minutes and then concentrated to dryness, some which gave diphenylmethyl-7-amino-3-methoxymethyl-3-cephem-4-carboxylate as a foam. This was dissolved in 20 ml of acetonitrile and in one portion added a stirred solution of 301 mg of D-N-tert.-butoxycarbonyl-2-naphthylglycine in 20 ml of acetonitrile containing 207 mg of N,N1-dicyclohexylcarbodiimide and 135 mg of hydroxybenzotriazole. The reaction mixture was stirred at 25° for 4 hours. It was then poured into 100 ml of ethyl acetate, after which the solution was washed once with 50 ml of aqueous sodium bicarbonate, once with 50 ml of 1N hydrochloric acid, once with water, then dried, after which the solvent was removed under reduced pressure to give diphenylmethyl-D-7-(N-tert.-butoxycarbonyl-2-naphthylglycylamido)-3-methoxymethyl-3-cephem-4-carboxylate as a white foam. The foam was dissolved in 2 ml of triethylsilane and 5 ml of trifluoroacetic acid, and the solution was stirred at 25° for nine minutes. Removal of the solvent by evaporation during redu-

pressure, yielded 260 mg of D-7-(2-naphthylglycylamido)-3-methoxymethyl-3-cephem-4-carboxylic acid trifluoroacetate salt.

The prepared salt was dissolved in 5 ml of water and 5 ml of acetonitrile, after which the pH of the mixture was adjusted to 4.5 with 1N ammonium hydroxide. The solution was lyophilized, where-

by a white solid was obtained, which was saturated by preparative reverse phase high pressure liquid chromatography, and a total of 20 mg of D-7-(2-naphthylglycylamido)-3-methoxymethyl-3-cephem-4-carboxylic acid was obtained.

NMR (DMS0-d6): <S 3.13 (s, 3H), δ 3.28 (q, 2H), δ 4.12 (s, 2H), δ 4.95 (d, 1H), δ 5 .65 (d, 1H), 67.41-4.23 (m, 7H).

Example 14

p-nitrobenzyl-7-/N-tert-butoxycarbonyl-(6-methoxy-2-naphthyl)glycylamido/-3-methyl-3-cephem-4-carboxylate.

A stirred solution of 66 2 mg (2 mM) of N-tert-butoxycarbonyl-(6-methoxy-2-naphthyl)glycine in 100 ml of acetonitrile containing 500 mg (2 mM) of N-ethoxycarbonyl-2-ethoxy-1, 2-dihydroquinoline, 770 mg (2>i2mM) p-nitrobenzyl-7-amino-3-methyl-3-cephem-4-carboxylate was added in one portion. The reaction mixture was stirred at 25° for 16 hours and then concentrated to dryness to give an oil. This was dissolved in 50 ml of ethyl acetate, after which the solution was washed with 25 ml of 1N hydrochloric acid, 25 ml of aqueous sodium bicarbonate and then with water. The solution was dried and concentrated to dryness, yielding 1.3 g of p-nitrobenzyl-7-/~N-tert-butoxycarbonyl-(6-methoxy-2-naphthyl)glycyl-amido7-3-methyl-3-cephem- 4-carboxylate.

NMR (CDCl 4 ). 61.39 (s, 9H), 6 2.08 and 2.15 (two singlets, 3H), 6 3.34 (q, 2H), 6 3.90 (s, 3H), 6 4.9 (m , 1H), 6 5.29

(s, 2H), 6 5.31 (s, 1H), 6 5.68 (m, 1H), 6 7.08-8.25 (m, 12).

Example 15 D-7-(6-Methoxy-2-naphthyl)glycylamido-3-methyl-3-cephem-4-carboxylic acid.

To a stirred suspension of 1.4 g of 5% palladium on carbon in 50 ml of ethanol was added in one portion 1.3 g of p-nitrobenzyl-7-/'<->N-tert-butoxycarbonyl-(6-methoxy-2-naphthyl )glycylamido/-3-methyl-3-cephem-4-carboxylate. The reaction mixture was stirred for 3 hours at 25°C under a hydrogen pressure of approx. 4.0 kg/cm<2>. The reaction mixture was then filtered, and the filter cake washed with fresh ethanol. The filtrate was concentrated to dryness by evaporation under reduced pressure to give 1.1 g of !-[_ N -tert-butoxycarbonyl-(6-methoxy-2-naphthyl)glycylamido7-3-methyl-3-cephem-4-carboxylic acid.

The acid produced was dissolved in 5 ml of trifluoroacetic acid,

and the solution was stirred at 25° for 5 minutes. 20 mL of water was then added, and the aqueous solution was lyophilized for 12 hours to give D,L-7-(6-methoxy-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid trifluoroacetate .; The salt produced was redissolved in water and purified by high pressure liquid chromatography to give D-7-(6-methoxy-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid.

NMR (TFA-d^): δ 2.42 (s, 3H), δ 3.50 (q, 2H), δ 4.30 (s, 3H), δ 5.39 (d, 1H), δ 5 .8-6.1 (m, 2H), 1 7.42-8.30 (m, 6H).

Example 16

By following the procedure described in examples 14 and 15, p-nitrobenzyl-7-amino-3-methyl-3-cephem-4-carboxylate was reacted with D,L-tert.-butoxycarbonyl-(6- hydroxy-2-naphthyl)

glycine in the presence of EEDQ, whereby p-

nitrobenzyl D, L-7-(/~N-tert.-butoxycarbonyl-(6-hydroxy-2-naphthyl)glycylamido/-3-methyl-3-cephem-4-carboxylate.

NMR (CDCl 3 ): δ 1.41 (s, 9H), δ 2.02 and 2.10 (two singlets,

3H), 6 2.9-3.5 (m, 2H), 6 4.9 (m, 1H), 6 5.23 (s, 2H), 6 5.35 (m, 1H), 6 5, 6-6.0 (m, 2H), 6 6.72-8.21 (m, 12H).

The compound produced was then reacted with hydrogen and

5% palladium on carbon, whereby D,L-7-/_ N-tert.-butoxycarbonyl-(6-hydroxy-2-naphthyl)glycylamido7~3-methyl-3-cephem-4-carboxylic acid was produced.

NMR (CDCl 3 ). 6 1.15 (s, 9H),6 1.82 and 1.89 (two singlets,

3H), 6 5.65 (two duplicates, 1H), 6 5.23 (m, 1H), 5 6.0

(m, 1H), δ 6.7-7.9 (m, 7H), δ 8.5 (m, 2H).

A reaction between the above compound and trifluoroacetic acid gives D,L-7-(6-hydroxy-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid. The isomers were separated by high pressure liquid chromatography, yielding D-7-(6-hydroxy-2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid.

Example 17 D-7-(2-Naphthylglycylamido)-3-cephem-4-carboxylic acid trifluoroacetate. A stirred suspension of 2.0 g (10 mM) 7-amino-3-cephem-4-carboxylic acid in 15 ml of acetonitrile was added in one portion to 8 ml of 30 mM bis-trimethylsilyl)trifluoroacetamide. The mixture was stirred at 25° for \ hour and then cooled to 0° and in one portion to

placed a stirred solution of 2.0 g of 6.6 mM D,L-N-tert.-butoxycarbonyl-2-naphthylglycine in 15 ml of acetonitrile containing 1.73 g (7.0 mM) of EEDQ. The reaction mixture was stirred for 1 hour at 25° and then concentrated to dryness to give an oil. This was dissolved in 100 ml of ethyl acetate, washed four times. times with 25 ml portions of IN hydrochloric acid, twice with sodium chloride solution and then dried. The solvent was removed by evaporation under reduced pressure to give a white

foam which was then dissolved in 25 ml of trifluoroacetic acid, where-

after the solution was treated with ultrasound for 5 minutes at 25°C. The reaction mixture was then concentrated to dryness and treated with diethyl ether to give 1.8 g (55% yield) of D,L-7-(2-naphthylglycylamido)-3-cephem-4-carboxylic acid trifluoroacetate. The product was chromatographed on reverse phase C^g silica gel, eluted with 8 L of a solution of 1% acetic acid and a gradient of 95% water -5% aceto-

nitrile to 85% water -15% acetonitrile (v/v). Appropriate fractions were combined, after which the solution was removed by lyophilization, yielding 157 mg of D-7-(2-naphthylglycylamido)-3-cephem-4-carboxylic acid.

IR (KBr): 1771.75 cm<-1>(3-lactam),

NMR (TFA-d^): 63.55, (q, 2H), δ 4.08 (s, 1H), δ 5.6-6.0

(m, 2H), 6 7.5-8.1 (m, 7H).

Example 18

α-Methoxyamino-α-(8-chloro-2-naphthyl)-acetamido-3-methyl-3-cephem-4-carboxylic acid.

420 mg of α-methoxyimino-α-(8-chloro-2-naphthyl)acetic acid was converted to the acid chloride by a reaction with excess chlorine and 500 mg of triphenylphosphite in 20 ml of dichloromethane. A stirred solution of 350 mg of 7-amino-3-methyl-3-cephem-4-carboxylic acid in 5 ml of dichloromethane containing 2 ml of bis(trimethylsilyl)trifluoroacetamide was added in one portion to the reaction mixture. The mixture was stirred at 25° for 6 hours, and diluted by adding 20 ml of methanol. The solvent was removed by evaporation under reduced pressure to give 7-[α-methoxyimino-α-(8-chloro-2-naphthyl)acetamido7-3-methyl-3-cephem-4-carboxylic acid. The product prepared was dissolved in formic acid containing zinc metal dust, which gave, after isolation and purification, 7-(8-chloro-2-naphthyl)glycylamido-3-methyl-3-cephem-4-carboxylic acid.

In a similar manner, the following compounds were prepared: 7-(8-nitro-2-naphthyl)glycylamido-3-chloro-3-cephem-4-carboxylic acid.

7-(8-hydroxy-2-naphthyl)glycylamido-3-methoxy-methyl-3-cephem-4-carboxylic acid,

7-(8-amino-2-naphthyl)glycylamido-3-methyl-3-cephem-4-carboxylic acid, and

7-(8-Methoxy-2-naphthyl)glycylamido-3-methyl-3-cephem-4-carboxylic acid.

Naphthylglycylcephalosporins provided by the present invention are valuable antibiotic substances

or intermediates for the production of such ."J Although the compounds are active in relation to the spectrum of gram-positive and gram-negative bacteria, they are particularly effective against a number of gram-positive bacteria. The antibiotic compounds are particularly valuable for the treatment of infections in animals and humans, which are caused by gram-positive microorganisms. The compound is particularly effective in the treatment of upper respiratory infections and similar diseases caused by severe influenza, S. aureus and S pyogenes, etc. The compounds are also effective in the treatment of diseases caused by anaerobic cocci, such as Peptostreptococcus anaerobius, Peptostrept. intermedius, Peptostrept. products, Peptococcus saccharolytius, P. prevotii, P. avaerobius, Propionbacterium acnes, Fusobacterium nacrophorum, etc.

A typical and preferred compound according to the present invention is 7-(2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid, i.e. the compound from example 3. The antibacterial activity of this compound and several other compounds according to the present invention, has been determined by a standard in vitro agar dilution test against a number of Gram-positive microorganisms. The following table indicates typical minimum inhibitory concentrations (MIC) in ug/ml for the mentioned compounds when tested against the indicated microorganisms. MICs for several known compounds are also listed for comparative purposes.

The above-mentioned data clearly show that compounds according to the present invention have a strong antibacterial activity.

In addition to having a strong antibacterial activity against a number of microorganisms, especially gram-positive organisms and anaerobic bacteria, compounds according to the present invention also have very favorable pharmacokinetic properties

creates in animals. When e.g. 7-(2-Naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid was administered to rats in an intravenous dose of 20 mg/kg, then the plasma concentration after 1 hour was 18.6 ug/ml, after 4 hours 14, 1 yg/ml, and after 2 4 hours the plasma concentration was still 1.96 yg/ml. The connection was

effectively orally absorbed in rodents and shows higher concentrations and longer duration of action than cefaclor and cefalexin.

The compound according to the present invention also has stability against B-lactamases. Table IV shows results from

comparative studies between several cephalosporins (lower numbers indicate greater stability against the specified lactamase).

The favorable pharmacokinetic properties of the present inventions, coupled with their excellent oral antibacterial activity and stability to 3-lactamase, make the compounds particularly valuable in the treatment of a variety of diseases caused by bacteria. The compounds are particularly well suited for treatment for outpatients, especially individuals suffering from mild respiratory infections caused by gram-positive microorganisms.

The present invention also provides methods for the treatment of animals and patients suffering from bacterial diseases, or who are suspected of developing a bacterial infection. The treatment method itself includes the introduction of an animal or a patient who may need treatment,

an antibacterially effective amount of a naphthylglycyl-cephalosporin antibiotic. The procedure can be practiced therapeutically or prophylactically. The amount of active antibiotic to be administered will depend on the particular compound used, the degree of disorder being treated, as well as the individual being subjected to treatment, and corresponding factors of the type encountered when it comes to such treatments. Normally, however, the compounds will be added in doses of from 0.5 to 50 mg/kg body weight, preferably from 1 to 10 mg/kg body weight. Such amounts can be added once or twice a day, depending on the need and the particular disease being treated. A typical daily dose for an average adult patient will be from 200 to approx. 500 mg/day.

The antibiotic compounds provided by

of the present invention are active both by oral and parenteral administration, and can consequently be prepared for a

each suitable delivery method. Such preparations are also included

in the present invention. The preparation according to the present invention will contain from 0.1 to 95% by weight of an active naphthylglycyl-cephalosporin antibiotic according to the present invention, mixed with a pharmaceutically acceptable carrier or diluent. Typical preparations will contain from approx. 10 to approx. 60% by weight active ingredient, preferably from 2 to 50%.

For convenient oral administration, the compounds can be formed

with a number of different known diluents or carriers and the type used for the production of oral preparations, and can optionally be processed into tablets, pills, encapsulated pills or gelatin capsules. Typical carry-

substances and diluents commonly used in the pharmaceutical industry for this purpose include potato starch, corn starch, sucrose, dextrose, microcrystalline cellulose, dicalcium phosphate, alginic acid, acacia; lubricants such as magnesium stearate; binders such as gum, tragacanth or gelatin, as well as flavorings such as peppermint oil, cherry or strawberry flavorings, wintergreen oil and the like. The compounds can also be processed in the form of syrups etc., where known diluents are used, such as fatty oils, methyl or propyl parabens, suitable dyes and flavourings. The compounds can also be prepared in the form of preparations for long-term regulated supply of the active ingredient over a longer period of time.

Antibiotics according to the present invention can also be prepared for parenteral administration, e.g. intravenously, intramuscularly or subcutaneously, as well as transdermally. Such preparations will normally contain from 0.1 to 20% by weight of active ingredients. For this purpose, you can use typical diluents and carriers, e.g. isotonic saline solutions, dilute aqueous dextrose, polyfunctional aliphatic, alcohols or mixtures of these, e.g. glycerol, propylene glycol or polyethylene glycol. Parenteral preparations may also contain preservatives such as phenethyl alcohol, methyl and propylparabens or timoro-

saddle. If necessary, from 0.05 to 0.2% by weight of an antioxidant such as sodium metabisulfite or sodium bisulfite can be added. For intravenous preparations, concentrations with up to 0.05 and up to 0.25 mg/ml of active ingredient will be used, and for intramuscular injection a preferred concentration of the active ingredient will be from 0.25 to 0.50 mg/ml.

The following examples illustrate typical preparations that can be used.

Example 19

Preparation of oral suspensions

The sorbitol solution was added to 40 ml of distilled water, after which naphthylglycylcephalosporin was added in suspended form. The saccharin, sodium benzoate and flavoring substances which were dissolved were then added. The volume was adjusted to 100 ml of distilled water. Each ml of the syrup contains 5 mg of the naphthylglycyl-cephalosporin antibiotic. This oral preparation is very suitable for pediatric use.

Example 20

Preparation of 250 mg capsule

The ingredients were mixed until they obtained a homogeneous mixture, after which the mixture was encapsulated in gelatin capsules. Such capsules can e.g. administered orally in a quantity of from one to two per day.

Example 21

Preparation of parenteral solution.

In a solution of 700 ml of propylene glycol and 200 ml of distilled water for injection, 20.0 g of D-7-(2-naphthylglycylamido)-3-methyl-3-cephem-4-carboxylic acid hydrochloride was dissolved.

The pH of the solution was adjusted to 5.5 with hydrochloric acid, after which distilled water was added to a volume of 1000 ml. The preparation was sterilized, filled into 5.0 ml ampoules each containing 2.0 ml (representing 40 mg of active ingredient), after which the ampoules were sealed under a nitrogen atmosphere.

Claims (20)

1. Method for producing compounds with formula I where R 7 and R 8 independently of each other are hydrogen, halogen, hydroxy, C 1 -C 4 -alkyl, C 1 -C 4 -alkoxy, nitro, amino, C 1 -C 4 - alkanoylamino, C, -C .-alkylsulfonylamino, or where R and R 8 together can form methylenedioxy, A and B are both hydrogen or can together form a double bond, R is hydrogen, an amino protecting group, hydroxy or 3 2 3 methoxy, and R is hydrogen, or R and R together can form the group: wherein M and L are independently C 1 -C 1 -alkyl, R 4 is hydrogen, methoxy or methylthio,-1-4 R is hydrogen, methoxy, methyl, halogen or methoxymethyl, R g is hydrogen, or a carboxy protecting group, provided that R <2> is only hydroxy or methoxy when A and B are a double bond, and that A and B are a double bond, and that A and B are both hydrogen, when R < 3> is different from hydrogen, or a pharmaceutically acceptable salt of such a compound, characterized in that one (A) acylates a compound of formula II with an acylating agent of formula III or an activated derivative of such a compound, where A, B, R<1> , R<2> , R <4> , R^ and R^ are as defined above, after which any amino- or carboxyl-protecting agent present is removed groups: 30 (B) deblocks a protected acid of formula I, wherein R yrs^ Qj^ K)g^ is a carboxy-protecting group, whereby a compound of formula I is provided-we-V- where R^, is hydrogen; (C) removes an optional amino-protecting group R from ij^ 2>5 a compound of formula I to thereby produce a pre- bond with formula I, where R 2 is hydrogen; (D) and when it is desired to produce a compound where R 2 and R 3 together form a group of formula : reacts a compound of formula I, where R 2 and R 3 are both hydrogen, with a ketone of formula M-C(0)-L where M and L are as defined above; or (E) reduces a compound of formula I where A and B together form a double bond and R 2 is hydroxy, or methoxy, thereby producing a compound of formula 2 In which A, B and R are hydrogen; and (F) if it is desired to form a salt of a compound of formula I, (G) or if desired, converting a salt of a compound of formula I to the free amine or the free acid. 2. Method for preparing a compound of formula I according to claim 1, characterized in that R 2 and R 3 together form the group or a pharmaceutically acceptable salt of such a compound. 3. Process for the preparation of a compound of formula I according to claim 1, characterized in that A and B together form a double bond, and R 2 is methoxy, or a pharmaceutically acceptable salt of such a compound. 4. Process for producing a compound of formula I according to each of claims 1 to 3, characterized in that R<1> is or a pharmaceutically acceptable salt of such a compound. 5. Process for producing a compound of formula I according to each of claims 1 to 3, characterized in that R <1> is or a pharmaceutically acceptable salt of such a compound. 6. Method for producing a compound of formula I according to each of claims 1 to 3, characterized in that R <1> is or a pharmaceutically acceptable salt of such a compound. 7. Process for the preparation of a compound of formula I according to each of claims 4 to 6, characterized 7 8 in that R or R independently of one another is hydrogen, halogen, hydroxy, C 1 -C 4 -alkoxy, micro or amine, or a pharmaceutically acceptable salt of such a compound. 8. Process for the preparation of a compound of formula I according to claim 7, characterized 7 8 in that R or R are independently hydrogen, halogen, methoxy or hydroxy; or a pharmaceutically acceptable salt of such a compound. 9. Process for the preparation of a compound of formula I, according to claim 7 or 8, characterized by 7 8 that R and R are both hydrogen; or a pharmaceutically acceptable salt of such a compound. 10. Method for producing a compound of formula I according to each of claims 1 to 9, characterized in that R 5 is methyl, chlorine, hydrogen, methoxymethyl or methoxy; or a pharmaceutically acceptable salt of such a compound. 11. Process for the preparation of a compound of formula I according to claim 10, characterized in that R<5> is methyl or chlorine; or a pharmaceutically acceptable salt of such a compound. 12. Process for preparing compounds of formula I according to any one of claims 1 to 11, characterized in that R 4 is hydrogen; or a pharmaceutically acceptable salt of such a compound. 13. Method for producing a compound of formula I according to each of claims 1 to 12, characterized in that R is hydrogen; or a pharmaceutically acceptable salt of such a compound. 14. Process according to claim 1, characterized in that !-/_ D-(2-naphthylglycylamido7-3-methyl-3-cephem-4-carboxylic acid, or a pharmaceutically acceptable salt of this compound is prepared. 15. Process according to claim 1, characterized in that 7-[D-(2-naphthylglycylamido Y)-3-methyl-3-cephem-4-carboxylic acid tetrahydrate is prepared. 16. Process according to claim 1, characterized in that 7-[D-(2-naphthylglycylamidoT7-3-methyl-3-cephem-4-carboxylic acid hydrochloride monohydrate) is prepared. 17. Method according to claim 1, characterized in that 7-[D-(2-naphthylglycylamido)-7-3-chloro-3-cephem-4-carboxylic acid is prepared, or a pharmaceutically acceptable salt of this compound. 18. Process according to claim 1, characterized in that one prepares 7-£~ D-(2-naphthylglycylamido)-7-3-methoxy-3-cephem-4-carboxylic acid, or a pharmaceutically acceptable salt thereof. 19. Compound with formula I or a pharmaceutically acceptable salt of such a compound, characterized by being produced by a method according to each of claims 1 to 18. 20. Compound of formula I according to claim 1, (25 characterized in that R <2> is an amino protecting group, or when A and B together form a double t bond^ ^ é^ T^ y^'^ ^which/is R2^ hydroxy or methoxy; or R<6> is a carboxy-I protecting group.