Improvements relative to derivatives of naphtylglycyl-cephalosporines.

2

7, 8

or R ' and R each independently represent 1¹ each other a hydrogen atom, a halogen atom, a hydroxy group, an alkyl group c ^ ^ - C., an alkoxy group in c ^ ^ - C., a nitro group, a group

an amino, an alkanoyl group (in C, c.) amino group, a group

4■■* η grams of alkyl (c ^ ^ - C.) sulfonylamino or when R ' and R are taken together, they form a group methylene-dioxyethylene $

A and b each represent a hydrogen atom or, taken together, they complete a double bond;

R represents a hydrogen atom, a

amino protecting group, a hydroxy group O

or methoxy and r° represents a HY -

2, 7

drogene or R and R, taken together, represent

Μ L.

/ Ν

/ VBE1

where Μ and L are each independently of one another alkyl group of C ^ ^ - C. Y

represents a hydrogen atom, a methoxy group or a methylthio group ;

The R~* represents a hydrogen atom, a methoxy group, a methyl group, a halogen atom or a methoxymethyl group J.

The R ^ represents a hydrogen atom or

a protecting group of carboxy group;

2

with the proviso that R is a hydroxy group or a methoxy group only when a and b complete a double bond and a and b

?

each hydrogen atom when R is other than

j is hydrogen

or a pharmaceutically acceptable salt thereof, is useful as an antibiotic active orally or as an intermediate for this antibiotic.

The preferred compounds provided by the present invention include those having the formula (I-) wherein

The R * represents

7, 8

while R^/ and R have the meanings defined

above. In this group, preferred compounds

2

include those in which R is a hydrogen atom, a protective group of the amino group, a hydroxy group or a methoxy group and R ^ is a hydrogen atom or a protective group of the carboxy group.

Another preferred group of compounds includes those in which R '*' represents

while ^ and R have the meanings defined above. Particularly preferred compounds falling within this group include those in which a, and b, the R ^, the R ^, the R ^ " ^ and R each represent a hydrogen atom.

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

wherein R ^, ^ ^ where R and R have the meanings defined above. The compounds far preferred are those in which R ' represents a hydrogen atom, a halogen atom, a hydroxy group or a methoxy group, the R ^ represents a methyl group or a chlorine atom and R ^ represents bydrogene, or the pharmaceutically acceptable salts of these compounds as sodium salts or potassium.

The present invention also provides a pharmaceutical formulation comprising a naphtylglycylamido-cephalosporin derivative of formula (I-) or a pharmaceutically acceptable salt thereof in association with a carrier, diluent or excipient for the derivative or salt. A preferred formulation is a formulation suitable for oral administration.

In another application of the present invention, there is provided a method for treating bacterial infections in animals, comprising administering, to the animal, an effective amount of an antibacterial compound of formula (I-) or a pharmaceutically acceptable salt thereof. In a preferred method of treatment, comprising administering the derivative dîenaphtylglycylamido-cephalosporin orally to treat diseases caused by gram-positive bacteria.

Further, there is provided a method for preparing a compound of formula (I in), the method comprising the steps of:

(Has) acylating a compound of formula (it):

with an acylating agent of formula (Ill-)

P S²

V

R¹ - C-C-0H

""

WITH 0

, - 12

or an activated derivative thereof, has, b., R., R., R., R.' ^ and R have the meanings defined below dessùs, (lll) 4

and then optionally removing either protecting group of the amino group or the carboxy group present the j

(D) releasing a protected acid of formula

(i-) wherein R ^ is a protective group of the carboxyl group to obtain a compound of formula (I-) wherein is hydrogen;

(D) removing a protective group of the

amino group, the R ^, of a compound of formula (I-) for

2 obtain a compound of formula (I-) wherein R is a hydrogen atom the j

(D.) when it is desired to provide a compound

2, 2

wherein R and R, taken together, form a group of the formula:

L-M.

the X /

J

reacting a compound of formula (I-) wherein R and are each r°'d * hydrogenated, with a ketone of formula:

M-C-THE L - (0)

wherein m and L have the meanings defined above J or

(i) reducing a compound of formula (I) in

wherein a and b are taken together to form a

2

double bond and R represents hydroxy or

a methoxy group, to obtain a compound of formula

2

(the I) where a, b and R each represent a hydrogen atom j and

(E) optionally salifying a compound of formula (I-),

(Grams) or optionally converting a salt of a compound of formula (I-) acid or free amine

In the above formulae, the R * represents a group 2-naphthyl group or a 2 a-tetrahydronaphthyl "

Naphthyl and tetrahydronaphthyl may

7, 8 be unsubstituted, for example, when R ' and R each represent a hydrogen atom j-they can

be monosubstituted in positions 1, 3, 4t St. 6T-

8

7 or 8, for example, when r° atom is a hydro -

7

gene and that R ' is other than hydrogen or $

7, 8 else they can be disubstituted when THE R ' and R are both different from hydrogen. Among the groups by which the cores naphthyl and tetrahydronaphthyl may be substituted, there are the hydroxy group, the alkyl groups in the Cj c ^, the alkoxy groups in the Cj c ^, halogen atoms, nitro group, the amino group, the alkanoyl groups (in the Cj c ^ unsubstituted alkyl amino and (c ^ ^ - C.) sulphonylamino.

The expression "C alkyl ^ - ^ the c" has the meaning thereof as are well known in the technique to designate straight carbon chains and branched lower alkyl groups such as methyl group, ethyl group, isopropyl group, n-propyl group, the group isobutyle and the group tertiary- butyl. Similarly, the expression "alkoxy group in c ^ - ^ the c" denotes lower alkyl groups bound to the naphthyl ring by an oxygen atom. Among the alkoxy groups specific c ^ ^ - C., there are methoxy, ethoxy, n-propoxy groups the, the group n-butoxy group and group isobutoxy.

The expression "halogen atom" encompasses the fluorine atom, a chlorine atom, bromine and iodine atom. A preferred halogen atom is a chlorine atom. The expression "alkanoyl group (c ^ ^ - C.) alkylamino" denotes an acyl radical of an acid alcanoi ' that lower naphthyl or tetrahydronaphthyl ring-bonded through a nitrogen atom * these groups include the group forinylamino, the acetylamino group and the group butyrylamino, the expression "alkyl group (in the Cj c ^) suifonylamino" denotes a group such as methylsulfonylamino group, the group éthylsulfonylamino and

the group n-butylsulfonylamino.

2

R represents a substituent on the nitrogen atom of the glycyl group and it may include a hydrogen atom and a protecting group of the amino group. The expression "amino protecting group" designates any substituent known in the technique that can be attached to a nitrogen atom of the amino group and which is easily removed when desired. These protecting groups are often used during the preparation of the compounds of the invention and are designed to reduce the risk of undesirable side reactions is the pro -

duisant due to the presence of an amino group

2 free # although those compounds wherein R is a protecting group, have biological activity, it is envisaged that the compounds most wished -

bdenoting biologically are those in the -

2

which R is a hydrogen atom. Therefore, the

2

compounds wherein R is a protecting group of the amino group, are primarily used as intermediates in the synthesis of compounds more preferred free amino.

The precise nature of the protective group of the amino group is not critical to the invention and which can be used any well-known protecting group. Specific protective groups of amino group is described by J.W. the Barton in "circuit protective groups in an in-Biologique joining technology", J. F. mcomy,, DE,, air plenum press assembly, York, Υ Ν.., 1973 * chapter 2 and by Greene in "circuit protective groups in an in-Biologique constraint", by John Wiley & sounds, York, Ν·Υ,, 98 ΐ ΐ, Chapter 7 · these two references are mentioned herein for reference in their teachings concerning the amino protective group, s at the amino protective group that is used most commonly, can encompass the alkanoyl groups and the groups haloalcanoyle ^ - ^ c-c-Q-groups such as the formyl, acetyl, chloroacetyl, dichloracetyl, propionyl, hexanoyl 10, 3, 3 a-diéthylhexanoyle or IF-to-chlorobutyryl J alkoxy groups (c ^ - ^ the Cj Jearbonyle and alkenyl groups (in ^) oxycarbonyl groups such as methoxycarbonyl, tertiary- butoxycarbonyl and allyloxycarbonyl J substituted aryl groups (in) 15 oxycarbonyl groups such as the groups benzyloxyearbonylearylalcényloxycarbonyle, 4-nitrobenxyloxycarbonyl cinnamoyloxycarbonyle 5 haloalkoxy groups and (c ^ - Cjcarbonyle group such as 2, 2, 2 a-trichloréthoxycarbonyle j of same as the

20 alkenyl groups and arylalkyl groups such as benzyl, phenethyl, trityl or allyl.

Among other amino protective group is employed usually, there are the enamines prepared by reacting the compound

2.5 free amino with a keto-ester such as methyl or ethyl acetoacetate.

In addition represents

it hydrogen or amino protecting group,

2

in the above formulas, R may be taken together with^0 to complete a ring system to form compounds of formula:

wherein R ^^- ,, The R **, where R ^, m and L have the meanings defined above. Examples of these compounds are the acetonides, especially those wherein m and L each represent a methyl group. The compounds are prepared by reacting a glycylamido-cephalosporin wherein R and R^J each represent a hydrogen atom, with a ketone such as acetone. These cyclic compounds are particularly useful as antibacterial agents long acting.

In the above formula, the R ^ represents a hydrogen atom, a cation of an addition salt such as an ammonium cation or an alkali metal, for example, lithium, sodium or potassium J or a protective group of the carboxy group. The expression "carboxyl protecting group" denotes groups well known in the technique and usually used to block or protect the carboxylic acid functionality of a cephalosporin during chemical reactions involving other functional seats in the molecule and which can be removed easily when desired by commonly known techniques such as hydrolysis or hydrogenolysis. Specific protective groups of the carboxy group that must use according to the invention, include those described by e, Haslam in "circuit protective groups in an in-Biologique joining technology", super, chapter 5, as well as by Greene in "circuit protective groups in an in-Biologique constraint", super, chapter 5, documents are cited here for reference, among the protecting groups of the carboxy group that used conventionally, there can be mentioned, for example, the alkyl groups in c ^ - ^ c-Q-such as the methyl group, the group tertiary- butyl and decyl group; halo-alkyl CJ Cjq group such as 2, 2, 2 a-trichloroethyl group and the 2 a-iodoethylisopropylcarbonate J C arylalkyl groups_C. . - C.₁ - such as the benzyl group, the group 4-methoxybenzyl, 4-nitrobenzyl group, diphenylmethyl group and the triphenylmethyl group $alkanoyl groups (c ^ - ^ C. gjoxyméthyle such as acetoxymethyl group or the group propionoxymethyl;

as well as other groups such as phenacyl group, the groups 4 to-halophénacyle, the allyl group, the dimethylallyl group, sorting groups (alkyl the Cj c ^ silyl such as trimethylsilyl group, as well as related clusters.

The naphtylglycyl - and tetrahydronaphthyl-glycyl cephalosporins provided by the present invention can be prepared by several methods one of which includes coupling a core 7"^hasminocéphalosporine of formula (it):

(II)

The nuclei of 7 a-aminocephalosporin that must be employed in the synthesis of compounds of the present invention, are well known in the technique and are readily obtainable by methods known in the technique. For example, there can be obtained the cores 3-halo cephalosporins by the methods described in the Patent of the United States of America no. 3.925 " 372. Can be prepared cores 3-methyl-cephalosporin by expanding the core of sulfoxides of penicillin with subsequent cleavage side chains or hydrogenation of 3 a-acetoxymethyl derivatives.

The expression "activated derivative" means a derivative of the carboxy function making the acylating agent of formula (Ill-) reactive coupling with a primary amino group to form the amido bond which connects the acyl side chain to the core. Suitable activated derivatives, methods for their preparation and their use as acylating agents for a primary amine are well known to the skilled person. Preferred are activated derivatives; (has) an acid halide such as chloride or bromide, or (b.) alkanoyloxy derivatives such as anhydrides a formyloxy or acetoxy mixed (e.g., Y in the list below represents KCHO or-OCOCH ^). Other methods to activate the carboxy function may involve the reaction of the carboxylic acid with a carbodiimide (e.g., n, n-'a-dicyclohexyl carbodiimide or n, n-' a-diisopropylcarbodiimide) to give an intermediate reagent which reacts in-situ with the group 7~^has mid^{the n} DEGREES. This is described in detail below.

Of mêmej reagents naphtylglycyletétrahydronaphtylglycyle and defined by formula (III-) are common knowledge and are prepared by adopting methods well known. Specific derivatives of naphtylglycyletétrahydronaphtylglycyle and that can be employed to prepare the

compounds of the inventionj correspond to formula IC -

2, 7 8

above in which R, the R ' and r° can have the following meanings:

^:ÿi. 4' receive

THE CD "•OF O

The CD Fvs)

Coupling a naphtylglycinetétrahydronaphtylglycine or derivative with 4 - 7^hasminocéphalosporine can be effected by adopting acylation of current techniques.

For example, may be reacted naphtylglycyle of an acylating agent, in particular, chloride or acid bromide or a derivative alkanoyloxy such as acetoxy or a formyloxy anhydrides, with a cephalosporin nucleus by adopting conditions

conventional acylation. During these reactions

2 acylating, it is generally preferable that R is a protective group of the amino group and that either a protective group of the carboxy group. These protecting groups serve to minimize side reactions as undesirable and enhances the solubility characteristics of the respective reactants.

The acylation reaction is generally performed by combining approximately equimolar amounts of an acylating agent of formula naphtylglycyle or tétrahydronaphtylglycyle (-Ill) (for example, an acid halide or an acid anhydride mixed.) with the core 7"^hasniinocéphalosporine, the acylation reaction is normally carried out in a mutual solvent such as benzene, chloroform, dichloromethane, toluene, n, n-dimethylformamide or 1' acetonitrile and is usually completed after a period of about 1 to about 12 hours when carried out at a temperature between about and about 60 °c -20, during the reaction, may be optionally used an approximately equimolar amount of a base such as pyridine, triethylamine, aniline or sodium carbonate as acid scavenger.

The product can be isolated from the reaction mixture by removing the reaction solvent, for example, by evaporation under reduced pressure and, if necessary, further purification may be effected by adopting routine techniques such as chromatography, crystallization, 1' solvent extraction and other related processes.

In another alternative preferred method for coupling a naphtylglycyletétrahydronaphtylglycyle or derivative with a 7 7"^hasminocéphalosporine to obtain compounds of the invention, there is used a coupling reagent such as those usually used in the synthesis of the peptides. Among the specific coupling reagents that can be used, there are carbodiimides such as n, n-' a-diéthylcarbodiimide, n, or n-diisopropylcarbodiimide and n, n dicyclohexyl carbodiimide (CCD) > carbonyl coupling reagents such as carbonyldiimidazole; isoxazolinium salts such as 3 '-sulfonate-ethyl 5' a-phénylisoxazolinium; as are quinoline compounds such as n-ethoxycarbonyl-2 ethoxy 1.2 above dihydroquinoline (EEDQ),

Is typically applied coupling a core 7^_a with a derivative of minocéphalosporinenaphtylglycyle or tétrahydronaphtylglycyle using a coupling reagent peptides by combining almost equimolar quantities of acid derivative 7~^has >^I ' innocent by-ceph-a 3 a-EM 4-carboxylic acid, a derivative and a coupling reagent naphtylglycine peptides according to the following:

where R *, where R ^, ^ ^ Rvj the R and R have the meanings defined above. Preferably, during such coupling reactions, R is a protective group of the amino group and R^2 is a hydrogen atom or a protective group of the carboxy group. Any protecting group can be subsequently removed by conventional methods to give the active antibiotic of the invention.

The coupling reaction is normally carried out in a mutual solvent such as dichloromethane, acetone, water, 1' acetonitrile, n, Ndiméthylformamide or chloroform and is usually completed when performed for about 10 to about 90 minutes at a temperature between about and about 60 °c -20. Longer reaction times are not detrimental and may optionally be adopted. The product, namely a naphtylglycyl - or tétrahydronaphtylglycyl-cephalosporin, is readily isolated simply by removing the reaction solvent, for example, by evaporation under reduced pressure. The product may be further purified by conventional methods such as other acid-basfe extraction, chromatography, a salt formation or the like.

In another method of preparing compounds of the invention, there is used a naphtyloxime of formula:

0

THE R - 1 ACTIVATED C-C-OH GROUP¹¹

the II

NR

active or a derivative thereof, wherein R has the meaning as defined above and R - 1 nts _T- 2 perceives a hydroxy group or a methoxy group. When R represents a hydroxy group, it is specifi - 2 quement protected with a group such as a trimethylsilyl group or by a group like protective of the hydroxy group. These derivatives of naphthyl-oxime can be coupled with a cephalosporin nucleus by either of the methods described above to obtain a compound of formula:

COOR in which R¹ , R.⁴ , R.³ and R^{the U} have the meanings defined above. These compounds are useful as intermediates because they are reduced easily by normal methods to give the compounds of the present invention preferred naphtylglycylamido. Further, the oximes of formula above wherein κ is a hydrogen atom, or a salt thereof are useful as antibiotics *

The compounds having a nitro group on the side chain can be tétrahydronaphtylglycylenaphtylglycyle or modified to form other compounds of the invention. For example, the nitro substituent can be reduced by conventional methods of reducing or hydrogenation to give the corresponding derivative naphtylglycyl-cephalosporin substituted with an amino group and the amino group may optionally be acylated by reaction with an anhydride or an alkanoyl halide in the Cj-C.₄ or an alkyl halide D * (c ^ - ^ C. Jsulfonyle to form the alkanoylamino - or - alkylsulfonylaminonaphtylglycylamidotétrahydronaphtylglycylamidocéphalosporines and corresponding of the invention.

Similarly, are prepared - KOM

2, 3

laid of the invention wherein R and R^J are taken together to form the group

L-M.

THE N/

/ VBE1

0

it

by reacting a ketone of formula m c-L with

2 o.

a compound of the invention wherein R and R^J each represent a hydrogen atom, generally in the presence of an acid such as methane-suifonic or the like. The cyclic compounds formed, for example, the acetonides preferred wherein m and L each represent a methyl group, are particularly useful as antibiotics for oral administration because they are effective over long periods of time.

Other compounds of the invention that is expected to be antibiotics to activity-par 2 ticulièrement long, are those in which R is an alkanoyl amino-group protecting group such as formyl or acetyl. Conveniently these compounds prepared by single -

reacting a centeredly naphtylglycylamido-cephalosporin

2

wherein R is hydrogen, with an acylating agent alkanoyl in c ^ - ^ Q-c is, for example, formyl chloride or acetic anhydride. It is anticipated that these products will not only n-acylated as antibiotics in themselves, but also as prodrugs in that they will be hydrolyzed in an animal system to form the derivative naphtylglycyletétrahydronaphtylglycyle or related.

Since the side chains and tétrahydronaphtylglycylenaphtylglycyle cephem antibiotics of the present invention contain an asymmetric carbon atom, for example, when a is a hydrogen atom, the compounds of the invention may exist in the form of optical isomers, including D and L isomers of the compounds of the invention can be used in the form of a mixture of, L for treating bacterial infections in animals or optionally separating optical isomers and use them individually.

Although both isomers are effective antibacterial agents, an isomer appears to be stronger than the other and it is the d-isomer, thereby constituting a preferred application of the invention.

The separation resolution of optical isomers can êti ' th provided by methods conventional product on the cephalosporin of the invention or on the side chain of naphtylglycinetétrahydronaphtylglycine or used as the starting material. Is generally performs the separation of optical isomers by chromatography in high yield, by enzymatic cleavage or by racemization or crystallization chemical,, a particularly preferred method for obtaining d-naphtylglycine comprises reacting the mixture d, L with the benzaldehyde 1 * acide and optically active tartaric acid according to the method described in United States Patent'd * 3•976 * 680®America

As stated above, the popcorn -

laid the preferred are those in which

2

R of the above formula is hydrogen® compounds, which are primary amines, are of a basic nature and readily form pharmaceutically acceptable salts by reaction with acids. These salts which are pharmaceutically acceptable, are preferred forms of salts used for treating bacterial infections. "Pharmaceutically acceptable" salts are salts useful in chemotherapy of warm-blooded animals.

Among the specific acids conventionally used to form salts, there are 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, methane sulfonic acid, oxalic acid or acid the para- toluene-sulfonic acid. The compounds of the invention wherein and each represent a hydrogen atom, readily form internal acid addition salt, especially the hermaphroditic.

The compounds of the invention generally found in the form of crystalline solids and are allowed to crystallize in ordinary solvents such qjje ethanol, water, n, n-dimethylformamide, the and the like acétqne. In addition, compounds in which is hydrogen, 4-carboxylic acids are acids•these compounds are acidic and they readily form salts with organic and inorganic bases. The pharmaceutically acceptable Hsels¹¹ used according to the invention also encompass these base addition salts.

The compounds of the present invention often crystallize in the form of a solvate or hydration and may be used in this form. By way of example, the present invention provides a crystalline composition of a material which is 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4~carboxylique to four water molecules. The crystals are large, dense and stable and they have easy at kneading and grinding to be adapted to a pharmaceutical formulation, in particular, solid dosage forms such as capsules filled and the like, prepared the tetrahydrate of the present invention by isolating the acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic (preferred product of the invention) of an aqueous medium.

This crystal has the remaining properties hereinafter of X-ray powder diffraction, as measured with a camera a powder-metallurgical of 114 * 6 mm containing a copper radiation filtered nickel 1.5405 has:

Gap;, relative intensities, i/1 13.29 0744

10.53 0.22

7, 0.44 HR

6.58 1.00

6.09 0.22

5.64 0.33

5.36 0.88

5.14 0.28

4.86 0.17

4.66 0.11

4.44 0.33

4.12 0.28

3.92 0.17

3.75 0.22

3.65 0.94

3.51 0.38

3.34 0.72

2.94 0.39

2.78 0.17

2.69 0.14

2.58 0.11

2.46 0.17

2.42 0.17

2.32 0.11

2.25 0.03

2.18 0.14

2.10 0.06

2.04 0.06

In particular, this compound can be prepared by reacting a acid addition salt of the acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3" cephem-to-4-carboxylic acid with a base such as 1' sodium hydroxide or triethylamine to form the corresponding zwitterionactivatingfactor and crystallizing in the water. For example, it can dissolve a salt as the salt of 1 * acide trifluoroacetic or hydrochloric acid salt in water or a mixture of water and an organic solvent such as acetone or 1' acetonitrile. Is added a base such as 1' aqueous ammonium hydroxide to adjust the pH to a value between about 3 and about 5" the precipitate formed is the tetrahydrate of the invention and the recrystallized easily in water. Alternatively, the compound may be prepared according to the present invention by isolating the product of the acylation of the acid 7 -^has in Mino-to-3-methyl 3 a-cephem-to-4-carboxylic (7 a-DCAA) of a water-containing solvent. For example, can be acylating the 7 a-DCAA typically in the form of a silylated derivative with a mixed anhydride or an acid halide of d-2 a-naphtylglycine n-protected as previously described.

In general, the acylation can be carried out in an organic solvent such as 1' acetonitrile. As soon as the acylation is completed, the protective groups can be eliminated by conventional methods and the solution can be diluted acid 7 - (d 2~naphtyiglycyl~amide) - 3 methyl-3 a-cephem-to-4-carboxylic acid with water so that it contains about 10 to 50% by volume of water, the pH of the solution being adjusted to a value of about 3 to about 5" the crystalline product formed is acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic to four water molecules.

This crystalline form is very stable for extended periods of time and is very well absorbed after oral administration.

This feature is somewhat surprising since the compound has only a minimum solubility in water. For example, this compound forms a saturated solution in water at 37 °c according to the following table:

jng/LCR

Another example of a hydrated form D.^, a compound of 1 * invention is the hydrochloride D.¹ acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-~céphem 3 and 4-carboxylic acid to a water molecule "

The crystals of this compound are large, dense and stable and they easily lend themselves to mixing and grinding treatment to be adapted to a pharmaceutical formulation, in particular, in solid dosage forms such as capsules filled and the like "prepared hydrochloride to a water molecule by crystallizing the acid 7" (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid in a medium of aqueous hydrochloric acid.

The crystalline hydrochloride to a water molecule has the remaining properties of the following X-ray powder diffraction, as measured with a ' camera of a powder-metallurgical 114.6 mm containing radiation CuI O

vRE are filtered at 1.5405 nickel has:

Gap, D. % Relative intensities, l/l ^ 15.64 1.00

7.90 0.57

6.61 0.24

6.22 0.05

5.74 0.24

5.54 0.12

5.11 0.29

4.75 0.10

4.47 0.14

4.23 0.19

4.10 0.17

3.93 0.71

3.85 0.12

3.71 0.33

3.49 0.07

3.35 0.12

3.27 0.14

3.15 0.10

3.08 0.07

3.03 0.07

2.93 0.10

2.78 0.10

2.69 0.12

2.61 0.02

2.53 0.10

2.47 0.10

2.37 0.10

2.30 0.02

2.24 0.02

2.17 0.0 5

The hydrochloride can be prepared to a water molecule provided by the present invention by reacting the acid 7 - (0" 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic, typically in the form of a solvate or crystalline tetrahydrate as previously described, with hydrochloric acid in water or in a solution of water and an organic solvent such as acetone or 1' acetonitrile. Using a sufficient amount of hydrochloric acid to maintain the pH of the solution below about 2, typically about 0.1 to about 0.8. The reaction is normally completed after a period of about 3 hours when there about is performed at a temperature of about 0 to about 60 °c * the precipitate formed is the hydrochloride to a water molecule is isolated easily by conventional means.

Alternatively, this compound can be prepared by isolating the product of the acylation of the acid 7" amino 3 methyl-3 a-cephem-a 4 a-carcar.boxylic (7 A-DCAA) of a solvent containing aqueous hydrochloric acid.

For example, can be acylating the 7" DCAA, typically in the form of a silylated derivative, with a mixed anhydride or an acid halide of d-2 a-naphtylglycine n-protected. The acylation can be generally performed in an organic solvent such as acetonitrile. As soon as the acylation is completed, the protective groups can be eliminated by conventional methods and solution can be diluted acid 7 "(d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-a 4"^because t-oxylic>with water so that it contains about 10 to 50% by volume of water, the pH of the solution being adjusted to a value of about 0.1 to about 0.8 by addition of hydrochloric acid. The crystalline product formed is the hydrochloride acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule.

The following list gives examples of specific classes of naphtylglycyl - and tetra-hydronaphtylglycyl cephalosporins, as well as specific compounds provided by the present invention:

Has " preferred compounds of formulae

1. The d * R is hydrogen or

salt cation;

7 8

a) the R ' each represent a r° and

hydrogen atom;

aLL. The R ^ represents a methyl group;

la2. The R ^ represents a chlorine atom;

la3. The R ^ is bromo;

the R ^ la4•represents a fluorine atom;

la5. The R~* represents an iodine atom;

laô. The R "* represents hydro -

gene;

I

la7. R^J represents methoxy;

la8. The R " represents methoxy - *

methyl;

7

b) R.^/ represents a hydrogen atom and

8

R° represents 6-methoxy;

lbl-. The R~* represents a methyl group;

lb2, the R~* represents a chlorine atom;

lb3. The R~* represents methoxy;

lb4 ^ R-represents a methoxymethyl group;

c) the R ' represents a hydrogen atom and

grams

R° represents 7 fluoro;

Herein. The R "* represents a methyl group;

the I '. 7•^) OF O

lc2, represents a chlorine atom %

lc3 the R '* represents a fluorine atom j.

lc4. The R~* represents methoxy;

the R lc5•'* represents a group

j. mêthoxyméthyle

η

d) the R ' represents the d * water "

grams

gene and R is 6 hydroxy;

iFL. The R ^ represents a methyl group;

ld2. The R ^ represents a chlorine atom;

ld3. The R "* represents methoxy;

ld4. represents

bromine;

7>

e) the R ' represents a group 4 a-niéthniéth.yle

8

and r°~6 represents chlorine;

lele ^ the R represents a methyl group;

le2. The R ^ represents methoxy;

1 ℮ 3~R-β * represents a chlorine atom;

pharmaceutically acceptable salt thereof.

2. Those of the formula:

and R represents a hydrogen atom or a cation

salt,

the n 8

has, the R ' r° and each represent a hydrogen atom;

2al. The R ^ represents methyl 3 C.

2a2. R^J 3 represents a chlorine atom

2a3, the R~* 3 2a4 represents methoxy, ~* wherein R represents a hydrogen atom

b., THE R? represents a hydrogen atom and

grams

7 R° represents a " hydroxyl 3 2bl, the R ^ represents methyl 3 2b2, the R~* represents a fluorine atom 3 C.

2b3, R.^J 3 2b4 represents an iodine atom, the R^3 2b5 represents methoxy, ~* wherein R represents a hydrogen atom and c, represents the R ^ 1-methoxy and R^8 represents a groupter -3 2cl butyl -, the R^3 2c2 represents a methyl group, a methoxy group represents the R^3 2c3, the R^3 2c4 represents a chlorine atom. R° represents methoxy -

methyl 3

7, 8 THE R²⁷ 3~chloro and r° represents 7 represents a group- isop levelsropoxy 3

* Represents the R~im-hydrogen atom, 5

2dl,

2d2. R^J represents methoxy -

methyl 3

C.

r represents a methoxy group 2d3•3

C.

2d4aR^J 3 2d5 represents a methyl group. The R^3 represents a chlorine atom or a pharmaceutically acceptable salt thereof.

B_has Those of the formula:

1. ^ The R represents a hydrogen atom the R * ^

represents methyl 3

7, 8

9, R.^/ and r° represent

2

hydrogen and R represents a group tertiary3 - butoxycarbonyl

aLL. The R ^ is P-nitrobenzyl 5

la2. The R ** represents 2, 2, 2 a-trichloroethyl 5

la3 ^ the R represents a trimethylsilyl group 5

la4. The R^3 phenacyl represents a group

7, 8

b.. R^/ r° and each represent a hydrogen atom * represents the R ^ tertiary- butyl .3

lbl-. R represents a group tertiary- butoxycarbonyl;

lb2. R represents an acetyl group j is 2 lb3. R is benzyloxycarbonyl or p-nitro - 2 ;

lb4 ^ represents a chloroacetyl group R. }

The R ' represents a hydrogen atom, wherein R represents a group 7, 8 7-methoxy, 2 - R recirculation has a group 2, 2, 2 a-trichloréthoxycarbonyleVBE1

Herein. R is benzyl or p-nitro - 6 ;

lc2. The R ^ represents a sodium cation VBE1 lc3. The R ^ represents a methyl group;

The R ^ represents methoxy, R." * represents a chlorine atom VBE1

The R ' and R 7, 8 are each hydrogen, R is formyl 2;

2al. R^{the U} 2, 2, 2 a-trichoréthyle represents the j

2a2. Wherein R represents a hydrogen atom ^ $2a3. The R ^ represents a sodium cation;

3. The R ^ is methylthio, R-^ and R each represent a HY - j-drogene grams

CPU * j. 3al represents a methyl group. R represents a group tertiary2 - butyloxy - carbonyl ;

3a2. represents a group the parathe j - nitrobenzyl

b.. The R ** represents a bromine atom $

3bl. R represents a group tertiary2 - butoxy-carbonyl - j.

3b 2 R.^{the U} the j represents a trimethylsilyl group

c., R.⁵ 3 3cl represents a methoxymethyl group. R represents a hydrogen atom the d * 3, 2 3c2. R is 2 - trimethyl silyl 3

3c3. The R ^ represents allyl 3

represents a methylthio group, wherein R represents a hydrogen atom ^ r ° 8 and represents 6-methoxy 3

The R^3 represents methoxy

4al. R represents a group tertiary2 - butoxy-carbonyl - 3

4a2. 3 b. represents a methyl group. The R~* 5 represents a chlorine atom

4b1. R represents a hydrogen atom 3, 2 4b2. R represents a 3, 2 4b3 phenacyl group. R° represents a group tertiary- butyl;

C those of formula:

wherein R ^ represents

the R A represents methyl 3^

aLL. R represents a group the paranitrobenzyl}

la2. represents a tert-butyl group

grams

la3. R^{the U} y represents a hydrogen atom la4. The R ^ represents a trimethylsilyl group }

b., the R^3 represents a fluorine atom

lbl-. R is methyl j-C., the R~* represents a methoxymethyl group.

Non-limiting examples below are given to illustrate the invention further.

Preparation 1

Preparation of the 2 a-naphtylglycine (also called acid a-amino-OC - (2 naphthyl) acetic).

During 20 hours, was heated, to 50 °c, a solution of 15.6 grams (0.1 meters) in 700 ml of 2 a-naphthaldehyde a mixture of water and ethanol to 50% containing 1437 grams (0.3 meters) of sodium cyanide and 38.4 grams (0.4 meters) of ammonium carbonate. The reaction mixture was cooled and has been concentrated to approximately 400 ml by evaporation under reduced pressure, the solution was then acidified to pH 2 by addition of concentrated hydrochloric acid. By filtration, the precipitated solid was collected formed, it was washed with diluted hydrochloric acid, and then it was dried to obtain 22.1 g of 4~(2-naphthyl) - 2.4 imidazolidine dione.

During 2 hours and a half, was heated at reflux a solution of 5 grams (22 mm) of 4 - (2 naphthyl) - 2.4 imidazolidine dione in 100 ml of aqueous sodium hydroxide at there % (v/v).

Then, the reaction mixture was filtered, it was cooled and it was washed with ethyl acetate.

Was then diluted aqueous solution with hydrochloric acid until a pH value of ôn 5" 1 and has been filtered to obtain the 2 a-naphtylglycine, is selectively repeating the reaction to obtain larger quantities of the product.

We purified a sample of 10 g of 2 a-naphtylglycine by dissolving it in 125 ml methanol containing 3" 9 ROI of acetyl chloride.

The reaction mixture was filtered, the filtrate was then diluted with 5 ml of aniline. By filtration, the precipitated product was collected and has been dried to obtain 7 g of 2 a-naphtylglycine. Melting point: 219 and 221°.

Preparation 2

Splitting the 2 a-naphtylglycine.

Reacted with a mixture of d - and L-2 a-naphtylglycine with has a-aminoethylbenzene optically pure in the presence of n, n-' a-dicyclohexylcarbodiimide to n - (I-phenylethyl) - α-amino-- (2 naphthyl) acetamide. After acid hydrolysis, the separation of the diastereoisomers by chromatography on silica gel gave d-2 a-naphtylglycine (gold -190 degrees + 3 degrees) and L-2 a-naphtylglycine (gold=+ 190° to + 3 degrees).

Preparation 3

Preparation of 6 a-methoxynaphth-to-2 yl glycine,

Has been transformed 2 a-bromo 6 a-napthalene derivative 2 lithio-by reacting with the 1.nbutyl lithium. Then, reacted with it 1' diethyl oxalate with the 2 lithio-to-6 a-methoxynaphthalene obtain has-keto 6-methoxy-naphth-a 2 a-ylacétate ethyl. Is reacted compound with hydroxylamine hydrochloride and sodium acetate to obtain 1 'a-hydroxyiminomethyl-6 a-methoxynaphth and 2 a-ylacétate ethyl. During 3 hours, at a temperature of 0 °c, a solution was stirred 17 "55 g of 1' oxime in 600 ml of methanol containing 5" 3 g of zinc dust metal and 135 ml aqueous formic acid to 50 $(space/space). After filtering the reaction mixture, the solvent is removed by evaporation to obtain 10.3 g of the ct-amino-- (6 a-methoxynaphth-to-2 yl) ethyl acetate.

Nuclear magnetic resonance spectrum (CDCl ^):

e 1.20 (t-, 3:00) 6> 2.15 (e, 2:00) J. 6 3.89 (e, 3:00) ;

S 4 * 15 (m-, 1:00) _} (f4, 72 (e, 1:00 ^ 6, 08 - 6, 75 (w, 6:00).

The hydrolysis of the ester formed by reacting with 1' sodium hydroxide in-has given the 6 a-methoxynaphth and 2 a-ylglycine.

Preparation 4

The production of nter -- butoxycarbonyl 2 a-naphtylglycine.

Has an agitated solution of 10 grams (50 mm) of 2 a-naphtylglycine (preparation 1) in 100 ml of sodium hydroxide in-, 50 ml of tetrahydrofuran was added, followed by 30 grams (140 mm) di - carbonatetertiary- butyl. The reaction mixture was stirred during 4 hours to 24 °c. The product was isolated by washing firstly three times the reaction mixture with portions of 50 ml of diethyl ether, the mixture was then acidified to pH 2 by addition of concentrated hydrochloric acid. Has been removed several times the aqueous acidic mixture with ethyl acetate, was then combined extracts, were found to be washed with water, dried and then has been ' removed the solvent by evaporation under reduced pressure to obtain 12.8 grams (yield: 85 $) n -tertiary- butoxycarbonyl 2 a-naphtylglycine.

Nuclear magnetic resonance spectrum (dimethyl-sulfoxide d ^): ^ 2, 5 (e, 9:00) 1^6, 85 (e, 1:00);

e 7 * 28 - 7.9 (m-, 7:00).

By following the general methods described in preparations 1 - 4, the following compounds were prepared:

nter -- butoxycarbonyl-- (6 methoxy 2-naphthyl) glycin

Nuclear magnetic resonance spectrum (CDCl ^):

S 1.2 and 1.4 (two broad singlets, 9:00) ;

5 * 4 e (broad singlet, 1:00) agno ^ 6, 7 (broad singlet, 1:00) -, e 7, 03 - 7, 8 (the m, 6:00);

n -tertiary- butoxycarbonyl-- (6-hydroxy 2-naphthyl) glycin

Nuclear magnetic resonance spectrum (CDCl ^):

e 1, 2 - 1, 4 (broad singlet, 9:00) $<^ 5, 3 - 5, 9 (two broad singlets, 1:00) ; ^ Τό, 9 - 8.5 (m-, 7:00);

n -tertiary- butoxycarbonyl-- (6 chloro-2-naphthyl) glycin

Nuclear magnetic resonance spectrum (CDCl ^):

cF of 1.15 (e, 9:00) J. e5, 3 - 5, 7 (ln, 1:00) J. 67, 3 - 8, 3 (m-, 8:00).

Preparation 5

Chloride hydrochloride of has-amino-- (2 naphthyl) acetyl.

Was bubbled hydrogen chloride through a cold solution (0 °c) of 5 grams (25 mm) in 150 ml of 2 a-naphtylglycine dichloromethane for 20 min. Then, the reaction mixture was stirred while adding, in a portion, 7.6 grams (38 mm) of phosphorus pentachloride and has been continued stirring at 0 and 10 °C for 2 hours.

The solution was filtered, dry and solvent is eliminated by evaporation under reduced pressure to obtain 5.2 gm (yield: % Of SL) chloride hydrochloride of has-amino 0c - (2 naphthyl) acetyl.

Infrared ray absorption spectrum (paste);

1795 cm "¹ .

Analysis: calculated for c ^ ^ ^ H-c-no.

Theory: LC 27.68

Found: LC 27.69.

Preparation 6

Enzymatic cleavage of d, L 2 a-naphtylglycine,

By following the general method described

in the Patent of the United States of America № 3•386,888, reacted with 19.8 g of D, L N-chloroacetyl 2 a-naphtylglycine with 4 g of amidohydrolase n-acyl-L-amino acid in a 1,250 ml potassium hydrogen phosphate buffer 0, 1 Μ, at pH 7.0, containing

5 x 10~^ m of cobalt chloride to 6 water molecules.

The reaction mixture was stirred during 2 hours at 37 °c. We precipitate L-2 a-naphtylglycine and she was removed by filtration. The filtrate was acidified to pH 2 by addition of hydrochloric acid In and we extract the mixture twice with 500 ml portions of ethyl acetate. Have been combined extracts, were found to be dried and the solvent is eliminated by evaporation under reduced pressure to obtain 9j005 g of d-n--chloroacetyl 2 a-naphtylglycine (yield: 91.5 $)"

Analysis: calculated for the Cj ^ ^ ^ IC H-no.

Theory: 60.55 C. > 4.36 5 N 5 * 04 HR

Found: C. 60.62 $hr 4 * 34 * n 4 * 76.

[<47 -^2l2 - DEGREES

Was washed L-2 a-naphtylglycine collected by filtration with a buffer of pH 7 * with water and finally with 1' normal hexane, then dry air to give 6.82 grams (yield: 95 $) of L-2 a-naphtylglycine,

Analysis: calculated for

Theory: c-j-hr 5.55 71.63 6 * 96 n-ii

Found: 69.85 C. } THE N 6.51 5.62 $HR

P-|>⁵ ] + 195.2 DEGREES.

Preparation 7

Acid 8-nitrate 2 a-naphthoic acid.

Acid 5-nitrate 2 a-naphthoic acid.

Has an agitated solution of 400 ml of concentrated nitric acid, at a temperature of 60 °c, was added, portionwise, 18 grams (0.11 μ) 2 a-naphthoic acid. The reaction mixture was heated to 70 °c during 2 hours, it was cooled and has been added to 200 g of ice. The precipitate was collected by filtration and dried to obtain has the 18.8 grams (yield: 77 $) of a mixture of 5-nitrate 2 a-naphthoic acid 8-nitrate 2 a-naphthoic. has been transformed mixture into ethyl ester by reaction with ethanol in the presence of sulfuric acid. It has crystallized 5 g of the mixture of ethyl esters in 20 ml of ethyl acetate to obtain 800 mg of 8-nitrate 2 a-naphthoate ethyl (melting point 120 °c e) and 1.9 g of 5-nitrate 2 a-naphthoate ethyl.

Preparation 8 8-amino 2 a-naphtylformiate ethyl.

A solution is hydrogenated 11.2 grams

of 8-nitrate 2 a-naphthoate ethyl (obtained as described in the preparation 7) in 100 ml of ethanol in the presence of palladium-on-carbon 5% " Is^has the reaction mixture filtered and eliminated the solvent from the filtrate to obtain the 8-amino 2 a-naphtylformiate ethyl.

Preparation 9 8-hydroxy 2 a-naphtylformiate ethyl.

During 10 min, to a cold solution (0 °c) and agitated-of 9.5 grams (44' millimeters) of 8-amino 2 a-naphtylformiate ethyl in 150 ml 6n sulfuric acid, was added dropwise a solution of 3>1 gm (45 mm) of sodium nitrite in 25 ml of water. The reaction mixture was stirred during

50 min, then it was added to a hot solution (90 °c) of 90 ml water in 10 ml of concentrated sulfuric acid. The reaction mixture was stirred for 10 minutes at 90 °c, then it was cooled and it was extracted with dichloromethane.

Have been combined extracts, were found to be washed with brine, dried and were found to have been removed the solvent to obtain, after chromatographic purification, 1.7 g of 8-hydroxy 2 a-naphtylformiate ethyl. Melting point 136 and 137° 9.ï,

By following the same general method, which has been reacted 14.9 g of 8-amino 2 a-naphtylformiate ethyl nitrite tertiary- butyl and copper chloride (lu) to obtain 11.5 g of 8-chloro 2 a-naphtylformiate ethyl.

Preparation 10 8-methoxy-2 a-naphtylformiate ethyl.

24 Hours during, at a temperature of 25 °c, was stirred solution of 216 mg of 8-hydroxy 2 a-naphtylformiate ethyl (preparation 9) in 15 ml of acetone containing six drops of dimethyl sulfate and 150 mg of potassium carbonate.

The reaction solvent was removed and the product was dissolved in ethyl acetate, it was washed with hydrochloric acid to % O and with brine, it was dried and has been concentrated to obtain 200 mg of 8-methoxy-2 a-naphtylformiate ethyl.

Preparation 11

Acid a-methoxyimino-has - (8-chloro 2-naphthyl) acetic.

Was added, in one portion, a suspension of 9.2 g of sodium hydride in 50 ml of n, to an agitated solution of Ndiméthylformamide 5.6 grams (24 mm) of 8-chloro 2 a-naphtylformiate ethyl 4.4 grams (36 mm) methyl methylthiomethyl sulfoxide in 10 ml of n, n-dimethylformamide. The reaction mixture was stirred during 4 hours at 25 °c, then it was concentrated to dryness. The product was dissolved in 250 ml of ethyl acetate and the solution was washed with hydrochloric acid to 5%, saturated sodium bicarbonate and brine. The solution was dried and the solvent is eliminated by evaporation to obtain 4>73 grams (yield: 63 $) of L-oxo L - (8-chloro 2-naphthyl) - 2 a-methylthio-2-methyl - sulfinyléthane. During 30 min, at a temperature of 65 °c, was stirred solution (10 mm) 3>12 grams of product into 150 ml of formic acid and 12 ml of acetic anhydride. To the reaction mixture, was added 856 mg (4 mm) of sodium periodate and has continued agitation during 15 minutes•has cooled the reaction mixture and it was concentrated to dryness, the product was dissolved in ethyl acetate, it was washed with Bi sodium carbonate and brine and eliminated the solvent to obtain 1.2 gm (yield: 46 $) methylthio-oxo-has - (8 Ch! oropharyngeal 2-naphthyl) acetate.

During 16 hours, at a temperature of 25 °c, was stirred a solution of 220 mg of the product obtained above in 15 ml of methanol and 15 ml of water containing 0.83 ml sodium hydroxide In and 70 mg of methoxyamine hydrochloride. The reaction mixture was acidified to pH 2 by addition of hydrochloric acid in vivo. The acidic solution was extracted with ethyl acetate which was dried and concentrated to obtain 170 mg a-méthoxyiminoa - (8-chloro 2-naphthyl) acetic.

By following the same general method, prepared the following compounds ii

acid dialkyl niéthoxyimino-a-OC - (8-nitrate 2-naphthyl) acetic acid;

' j of the acid a-methoxyimino-has~(8-amino 2-naphthyl) acetic $

1¹ acid a-methoxyimino-has - (8-hydroxy 2-naphthyl) acetic therein and

acid-OC-methoxyimino-has - (8-methoxy-2-naphthyl) acetic.

Preparation 12

Preparation of 7 -^has in Mino-to-3-chloro 3 a-cephem-a 4" carboxylic.

In the Patent of the United States of America no. 3.925 · 372, there is described the synthesis of 7" phénylglycyland. oD and 3 chloro-~céphem 3 and 4-carboxylic currently known under the generic name "cefaclor". The reaction cefaclor with phosphorus pentachloride, methanol and water under conditions known for cleavage of side chains of cephalosporins gives acid 7 -^has mid^{the n} o 3-chloro 3 a-cephem-a 4 a-c^has the I ' boxylic.

In the same way, they can be prepared in the following cores for use in the synthesis of compounds of the present invention:

1' acid 7"^has in Mino-to-7 ~méthoxy and 3 a-bromo 3 a-cephem-to-4-carboxylic J.

7 acid "^has in Mino-a 3 a-cephem-to-4~carboxylique $acid 7^_am; the I -^no. "3" niéthoxyméthyl and 3 a-cephem-to-4-carboxylic J.

1•acid 7"^has "¹ the I^no. " 7^_niéthylthio-to-3-methyl 3~cephem-a 4 a-osrboxylic.

Preparation 13 The n, n-dimethylformamide acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3' a-cephem-a 4" - carboxylic a water molecule.

Has a suspension cold (-20 to -30 °c) and agitated 51 grams the sodium salt of d-[the n - (1-methoxycarbonyl) - 2 a-propenyl] and 2 a-naphtylglycine produced as by reacting 1' methyl acetoacetate with d-2 a-naphtylglycine, in 500 ml acetonitrile containing 250 ml * d-n, n-dimethylformamide, was added successively 0.44 ml of methane-suifonic, 0.45 ml of n * N-dimethylbenzylamine and 12.35 ml of methyl chloroformate. The reaction mixture was stirred during 2 hours at -20 to -30 °c after the addition. The reaction mixture was diluted by the addition of a cold solution (0 °c) of 34 * 24 g acid 7~^has in Mino-to-3-methyl 3 "cephem-a 4" C.^has * i-boxylic in 230 ml of acetonitrile containing 59 * 4 ml of n-methyl-monotriméthylsilyl-to-trifluoroacetamide. The reaction mixture was stirred during 2 hours at -20 to -30 °c, then it was heated to 0 °c. The reaction mixture was acidified by addition of l60 ml of hydrochloric acid is stirred in and the acid mixture while adding, in a portion, 17 * 85 g of semicarbazide hydrochloride, we adjusted the pH of the reaction mixture to 3 and has been kept by addition of triethylamine. After heating the mixture to 25 °c, it was filtered through a filter aid "through hyflo". The filtrate was diluted by addition of triethylamine to pH 6.2 and has been cooled to 0 °c for 45 min, then it was filtered. Was washed four times the filter cake with 50 ml portions of acetonitrile and it was dried at 35 °c during 7 hours under reduced pressure to obtain 6é^J , 8 g of N, N-dimethyl formamide acid 7" (D.^{, -} 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule. Yield: 86%, Karl Fischer titration: $4 * 69" nuclear magnetic resonance spectrum (trifluoroacetic acid): signals to 0 2.1 (E * 3:00),

3.2 (E, 3:00), 3.3 (ABq, 2:00), 3.4 (e, 3:00), 5.2 (1:00'd *) 45

Λ 7 // U-O-U- the L

5.8 (hD, 1:00), 5.8 (e, 1:00), 7, 3 - 8, 2 (m-, 7:00),

8.3 (e, 1:00).

Example 1

7 Acid trifluoroacetate salt (2 a-naphtylglycylamido) - - 3 methyl-3 a-cephem-to-4-carboxylic acid.

A stirred suspension of 1 has a g (4.7 mm) acid 7 -^has in Mino-to-3-methyl 3 a-cephem-to-4-carboxylic (7 a-DCAA) in 25 ml of âcétonitrile, is added, in one portion, 3.7 ml (14 mm) of bis - (trimethylsilyl) trifluoroacetamide. The reaction mixture was stirred at room temperature until all the solids dissolved, thereby indicating the complete formation of the Trimethylsilylamino of 7 a-DCAA.

In a separate flask, during 15 min, was stirred, at room temperature, a solution of 1.35 g of n - (4.5 mm)tertiary- butoxycarbonyl 2 a-naphtylglycine (preparation 4) in 20 ml of acetonitrile containing 1.1 grams (4.5 mm) of n-ethoxycarbonyl 2-ethoxy-L-, 2 a-dihydroquinoline (EEDQ). Then, this solution was added in one portion to the solution cold (0 °c) of âcétonitrile containing the Trimethylsilylamino of 7 a-DCAA obtained above. The reaction mixture was stirred for one hour at 0 °c, then it was heated to room temperature.

The solvent is eliminated by evaporation under reduced pressure to obtain an oil which was dissolved in ethyl acetate, which was washed twice with hydrochloric acid In and which was dried, the solvent was then removed by evaporation to obtain the acid 7 - (n--tertiary- butoxycarbonyl 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid as a foam.

Having dissolved the naphtylglycyl-cephalosporin n-protected resulting in 5 ml of trifluoroacetic acid, followed by trifluoroacetic acid is removed by evaporation under reduced pressure to obtain, after precipitation in diethyl ether 1 *, 5>7 g of the trifluoroacetate salt of acid 7" (2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-a 4 a-carboxylic acid.

Example 2

7 Acid "(2 a-naphtylglycylamido) -3" methyl-3" cephem-to-4-carboxylic to four water molecules.

Was heated a mixture of 5" 7 g of the trifluoroacetic acid addition salt of 1' example 1 in 55 ml of acetonitrile and water to 10 $(by volume/volume in) to about 50 °c, then it was filtered to remove undissolved solids. Then, the filtrate is diluted with 1' ammonium hydroxide to a pH of 1, 8m 4j5 * by filtration, the precipitate formed was collected and has been dried to obtain 3jl5 grams (yield: 72%) 7 acid^- (2 a-naphtylglycylamido) - 3 - methyl-3 a-cephem-to-4-carboxylic to four water molecules.

Example 3

Acid 7 - (d 2 a-naphtylglycylamido) - 3 a-inéthyl and 3 a-cephem-a 4 - carboxylic four water molecules.

We repeated the process of example 1

using 5 g of d-n-tert-butoxycarbonyl-2 a-naphtylglycine optically active and 5.6 g of 7 a-DCAA to obtain, after removing the N-protecting group, 6.8 grams (yield: 80 $) of the trifluoroacetate salt of d-7 - (2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid. Having dissolved the salt so formed in 90 ml of acetonitrile and 10 ml of water containing 5 ml of triethylamine. The reaction mixture was stirred for 20 minutes at 25 °c to and has been filtered. The filtrate was concentrated to dryness and the product was crystallized in water to give 2.9 g of acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4 carboxy - lic to four water molecules. Melting point calculation for ^ ^ ^ C. NgOgS

171 l80 °C (decomposition).

Analytical

Theory: 51.16 C.; H-n-j-5.80 8.95; e 6.93 found: 52.52 C. } 5.47 HR the I THE N 8.73 J E 6.83.

Nuclear magnetic resonance spectrum (dimethyl-sulfoxide d ^) 6 vBE1 1.9 (e, 3:00) j.</4, 8 (e, 1:00) J.

cT-4.9 (HD, 1:00) j.</5, 6 (HD, 1:00) I to 7, 49 - 7, 99 (m-, 7:00).

Example 4

7 Acid " (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic to four water molecules.

It has protected the sodium salt of d-2 a-naphtylglycine form an enamine by reaction with methyl acetoacetate. It has cooled a suspension of 102 grams (317.7 mm) of the sodium salt of d-2 a-naphtylglycine protected in 1.000 ml acetonitrile and 500 ml of n, n-dimethylformamide to -30 °c and has been stirred while adding, in a portion, 0.88 ml of methane-suifonic, was then added 0.90 ml of n, ~n-dimethylbenzylamine and 24.7 ml of methyl chloroformate. The reaction mixture was stirred during 2 hours to -30 °c, then it was diluted by dropwise addition of a solution of 68.5 grams (302.8 mm) acid 7-amino 3 methyl-3 a-cephem-to-4-carboxylic in 460 ml of acetonitrile containing 118.8 ml of hexamethyldisilazane. After the addition complete, the reaction mixture was stirred for about 2 hours to -30 °c, then it was heated to 0 °c. The reaction mixture was diluted by addition of 320 ml of hydrochloric acid in vivo, was then added 35.7 g of semd semicarbazide hydrochloride.

Was added 1' - ammonium hydroxide to adjust and maintain the pH to 3 while heating the mixture to 22 °c, is diluted further the reaction mixture by addition of 430 ml of water, then it was discolored by shaking for 15 minutes with 10 g of coal.

The reaction mixture was filtered through a filter aid "through hyflo" and has heated the filtrate to 40 °c. It has adjusted the pH to 4 by addition of ammonium hydroxide in vitro, after which the crystallization has started. Which comprises crystallizing is continued for about 30 min, was then carried the pH to 5.2 by addition of ammonium hydroxide in vivo.

The mixture was cooled to 20 °c, it was agitated for one hour and has been filtered " was washed twice the filter cake with portions of 50 ml of water and it was air dried to obtain 110.8 g acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic to four water molecules.

Analysis: the n ^ ^ calculated for the O ^ ^ 0 s at 4.

Theory: 51.16 C.; H-j-n-8.97 5.80 " E 6.83 found: 50.31 C. } H-n-j-5.62 8.87 } E 6.89 " Karl Fischer water analysis of:

Theory: 15.3 $

Found: 13.73 $

Nuclear magnetic resonance spectrum (trifluoroacetic acid) 2.2 (e, 3:00) 3, 3.21 (Q-, 2:00) 3, 5.08 (d-, 1:00) 3, 5.68 (m-, 2:00) 3 7, 4 - 8, 3 U-, 7:00) 3, 10.5 (e, 12:00).

Absorption spectrum infrared rays (on KBr);

I766, 1751, i696 cm."¹ .

Absorption spectrum ultraviolet rays (ch-OH-^) % λ 227, OOO £78.

λ_max. 265, £12,000.

Titration (66 $the n, n-dimethylformamide/water)

pKa of 5.5, 7.5, 11.2.

Example 5

7 - Acid hydrochloride (B 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule.

Has an agitated solution of 200 ml of water containing 10 ml 12n hydrochloric acid, is added, in one portion, 20 g of the solvate of n, n-dimethylformamide acid 7 - (Û-to-2-naphthylmethyl glycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid (preparation 13)" resized the reaction mixture at a pH of from 0.6 * d-by adding hydrochloric acid 12n, and then it was stirred at 25 °c for 45 mins. By filtration, the precipitated solid was collected and it was washed once with 50 ml of water. The crystalline material was dried to 35 and 40 °C for 16 hours to obtain 17.69 grams (92 $) 7 - acid hydrochloride (D~2-to-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule.

Analysis: ^ ^ c2qh22c1n calculated for 0 sec.

Theory: C. 53.15 ; 4.91 HR; THE N 9.30; 7.09 5 SEC.

0, 17.70 ; LC 7.85

Found: 52.75 C. ; H-n-j-4.85 9.85; 7.03 e;

0, 17.07 ii LC 7.60.

Karl Fischer titration:

Theory: 4.29 $

Found: 4.62 $.

Titration chlorides: theory: 7.85 j found: 8.16 and 8.02.

Absorption spectrum infrared rays (on KBr);

3040, 2940, 1768, 1707, 1533, 1232 cm "¹ .

Absorption spectrum ultraviolet rays (EfcOH)

λ 225, T- =100,000.

max.'

Titration (66 $the n, n-dimethylformamide/water, the volume/volume of) 5.5, 7.3.

Example 6

7 - Acid hydrochloride (d 2 a-naphtylglycylamido) - 3 a-niéthyl and 3 a-cephem-to-4-carboxylic acid to a water molecule.

Was heated, to 40 °c, a solution of

5 grams (10.6 mm) acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-céphe." n 4-carboxylic four water molecules (example 3) in 50 ml of acetone, then

the U

the acidified by addition of 1 ml with hydrochloric acid 12n. The mixture was stirred and has been diluted by adding 5 ml of water and ammonium hydroxide to a pH of 3.3 ôn " has a lowered the pH to 0.20 by dropwise addition of hydrochloric acid 12n, was then stirred the reaction mixture for 2 hours at 25 °c₀ By filtration, the precipitated solid was collected, washed it was twice with 10 ml portions of water and has been dried at 25 °c under vacuum for 2 hours to obtain 1,929 grams (yield: 4 - 0, 5 $) 7 - acid hydrochloride (d 2 a-naphtylglycylamido) - 3 a-méthy1 and 3 a-cëphem-to-4-carboxylic crystalline to a water molecule.

Chloride analysis; theory: 7.85 $j found:

7.96 $.

Example 7

Acid hydrochloride THE J -(D 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule.

Has a hot solution (40 °c) and agitated for 1 ml of hydrochloric acid 12n in 25 ml water and 25 ml of acetone, was added, in one portion, 5g acid 7 - (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-a 4 a-cai'cai' boxylic to four water molecules (example 3). An additional amount added 12n hydrochloric acid to the reaction mixture to adjust the pH to 0.15, the mixture was then cooled to 25 °c and it was stirred for 2 hours. By filtration, the precipitated solid was collected, washed with three seen by 10 ml portions of water and dried for 4 hours with the 25 °c to vacuum. It has been determined that the product was formed of 3,596 grams (yield: 75 $) 7 - acid hydrochloride (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule.

Karl Fischer titration:

Theory: 4 * 29%

Found: 5.95%.

Chloride analysis: theory: * 7 * 85%

found: 8.09%.

Example 8

Acid hydrochloride 7" (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid to a water molecule.

Was stirred, at 40 °c, a solution of 10 grams (21.3 mm) acid 7 "(d 2 a-naphtylglycylamido) -3" methyl-3 a-cephem-to-4-carboxylic to four molecules of water in 100 ml of water containing 5 ml of hydrochloric acid 12n, while adding dropwise 12n hydrochloric acid to a pH of 0.6. The reaction mixture was cooled to 25 °c and it was stirred at this temperature for one hour. The crystalline product was collected by filtration, washed it was twice with 20 ml portions of fresh water and has been dried for 3 hours at 25 °c under reduced pressure. It has been identified that the dried product was formed of 8.16 grams (yield: 84.5%) 7 - acid hydrochloride (d 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic crystalline lens to a water molecule.

Karl Fischer analysis:

Theory: 4 * 29%

Found: 5 * 37%.

Chloride analysis: theory: 7 * 85%

found: $7 * 62.

Example 9 7 - (N-tert-butoxycarbonyl 2 a-naphtylglycylamido) - 3 chloro-3 a-cephem-a 4 a-earboxylate P-nitrobenzyl.

Was added, in one portion, a solution of 260 mg of EEDQ in, the d * 50 ml acetonitrile containing 301 mg of n -tertiary- butoxycarbonyl 2 a-naphtylglycine to a cold solution (0 °c) and agitated 682 mg of 7-amino 3-chloro 3 a-céphen " - 4 carboxylate of P-nitrobenzyl in 50 ml of acetonitrile. The reaction mixture was stirred for 90 minutes at 0 °c, then it was heated to room temperature. The reaction solvent is eliminated by evaporation under reduced pressure to obtain the product as an oil. This oil was dissolved in ethyl acetate and gold. was washed with diluted hydrochloric acid, a buffer with a pH of 7 and finally, with the brine. After drying of the organic solution, the solvent is removed by evaporation to obtain 598 mg (yield 93 $ii) of 7 - (n-tert-butoxycarbonyl 2 a-naphtylglycylamido) - 3 chloro-3 a-cephem-to-4 carboxylate of P-nitrobenzyl.

Nuclear magnetic resonance spectrum (CDCl ^) ii ($1.21 (e, 9:00) Y ($3 * 1 - 3,8 (m-, 2:00) ;℮ L, 9 (two doublets, 1:00) J. 6 5.28 (e, 2:00) j.<^ 5, 4 - 6, 2 (m-, 3:00);

7 /, ' 2 - 8, 21 (the m, 12:00).

Example 10

Trifluoroacetate acid 7 - (2 a-naphtylglycylamido) - 3 chloro-3 a-cephem-to-4-carboxylic acid.

To a solution of 598 mg of 7~(d -tertiary- butoxycarbonyl 2 a-naphtylglycylamido) - 3 chloro-3~cephem-to-4 carboxylate of P-nitrobenzyl (example 9) in 50 ml of tetrahydrofuran containing 10 ml of methanol and 5 ml of ethanol, was added, portionwise, 1.20 g of palladium-on-carbon 5 $" was stirred the reaction mixture for one hour in a balloon hydrogenation of ATR policy under an initial hydrogen 4j06 kg/cm2.

Then, the reaction mixture was filtered and the filtrate solvent is eliminated by evaporation under reduced pressure to obtain an oil. This oil was dissolved in a buffer with a pH of 7 and resized the pH to 7 * 75 by addition of dilute sodium hydroxide. The aqueous solution was washed with 1 * acétate ethyl and diethyl ether, then it was acidified to pH 2.3 inches by adding hydrochloric acid. The acidic solution was extracted with ethyl acetate which was then dried and the solvent is eliminated by evaporation to obtain 360 mg (yield: 77%) acid 7 - (n--tertiary- butoxycarbonyl 2 a-naphtylglycylamido) - - 3 - 3-chloro-céphein-to-4-carboxylic acid.

Having dissolved acid thus formed in

6 ml of trifluoroacetic acid was stirred and the solution for 5 minutes at 25 °c. Was then adjusted the pH to 3>9 by addition of ammonium hydroxide diluted and eliminated the solvents by evaporation to obtain an oil. This oil was subjected to chromatography gradient on silica gel by eluting with reversed phase 0 - 25% acetonitrile, 1% acetic acid and 99 - 74% water (by volume/volume of).

Appropriate fractions were combined and the solvent was removed by lyophilization to give 49 mg of acid L 7 - (2 a-naphtylglycylamido) - 3 chloro-3 a-cephem-a 4 - carboxylic and 49 mg of d-7 - (2 a-naphtylglycylamido) - 3 chloro-3 a-céphein-to-4-carboxylic acid.

Nuclear magnetic resonance spectrum (trifluoroacetic acid a-d ^): 3 SEC.)% 2 (a Q, 2:00) ^ $5 * 49 (D., LHRH) J. 6 5, 8 - 6, 1 (m-, 2:00) J. 6 7 * 6 - 8,35 (m-, 7:00).

Example 11

Trifluoroacetate acid 7 - (2 a-naphtylglycylamido) - 3 methoxy 3 a-cephem-to-4-carboxylic acid.

During 30 min, to 25 °c, was stirred a solution of 1.05 g of D, L-nter -- butoxycarbonyl 2 a-naphtylglycine in 15 ml of acetonitrile containing 1.06 g of EEDQ, then it was added in one portion to an agitated solution of 1 g of acid 7 -^has in Mino-to-3-methoxy-3 a-cephem-to-4-carboxylic acid in 15 ml of acetonitrile containing 1 ml of bis (trimethylsilyl) acetamide - trifluoro. The reaction mixture was stirred during 2 hours at 25" C., then the I^O has concentrated to dryness by evaporation of the solvent under reduced pressure and having obtained an oil. This oil was dissolved in 10 ml of trifluoroacetic acid and maintained at the resulting solution to 0 °c during 5 min. By evaporating the solvent and purifying the product by chromatography on silica gel, the trifluoroacetate was obtained 89.5 mg of d, L 7 - (2 a-naphtylglycylamido) - 3 methoxy 3 a-cephem-to-4-carboxylic acid.

Absorption spectrum infrared rays (on KBr):

""* 1762.10 cm ^ - lactam).

Nuclear magnetic resonance spectrum (trifluoroacetic acid a-d ^): ^ 3, 2 - 4, 25 (w, 5:00) j.</5, 2 - 5, 9 (RNs, 3:00); 01,5 - 8, 3 (m-, 7:00).

Example 12

D-7 - (6 a-Chloronaphth and 2 a-ylglycylamido) - 3 methyl 3 - cephem-to-4-carboxylic acid.

Which was acylated with the 2-chloronaphthalene chloroxalate ethyl to form acid ethyl-α-ketoacid has - (6 a-Chloronaphth-to-2 yl) acetic. The reaction of this compound with 1' hydroxyl amine, followed by reduction and hydrolysis, gave the 6 a-Chloronaphth and 2 a-ylglycine. has been transformed by the latter in protected derivative on the n -tertiarybutoxycarbonyl.

To a solution stirred cold (0 °c) and

of 523 mg (1.5 mm) of 7-amino 3 methyl-3 a-cephem-a 4 - carboxylate in 300 ml p-nitrobenzyl acetonitrile, was added a solution of 500 mg (L-, 5 mm) of n -tertiary- butoxycarbonyl 6 a-Chloronaphth and 2 a-ylglycine in 100 ml of acetonitrile containing 369 mg of EEDQ.

The reaction mixture was stirred for one hour at 0 °c, then it was heated to room temperature and it was stirred for 48 hours additional.

Then, the reaction solvent is eliminated by evaporation under reduced pressure to obtain the product as an oil. This oil was dissolved in 100 ml of ethyl acetate and it was washed with hydrochloric acid in-, aqueous sodium bicarbonate and water, and then dry. By removing the solvent by evaporation, 770 mg was obtained a white solid (yield: 77%) of 7 - (Ν -tertiary- butoxycarbonyl 6 a-Chloronaphth and 2 a-ylglycylamido) - 3 methyl-3 a-cephem-to-4 carboxylate of P-nitrobenzyl.

Nuclear magnetic resonance spectrum (CDCl ^):

S 1.40 (e, 9:00); 2.12 FS (two singlets, 3:00) j.<^ 3, 40 (Q-, 2:00); S4 68 and 4>90 (two doublets, 1:00) J. 05.30 (e wide, 3:00) J. <F. 5j6 above 6.0 (m-, 1:00) J.

67, 2 - 8, 3 (m-, 8:00).

Eliminated the group P-nitrobenzyl protecting the carboxy group by hydrogenation of 770 mg of the compound obtained above with 1 g of palladium-on-carbon 5% in 50 ml of methanol containing 20 ml of ethanol under an initial hydrogen pressure of 3 * 85 kg/cm2. After completion the reaction after 55 min, the reaction mixture was filtered and eliminated the solvent of the filtrate to obtain an oil. This oil was dissolved in 50 ml of ethyl acetate containing a buffer of pH 7, was then acidified the aqueous layer to pH 2.3 with hydrochloric acid in vivo. The product was extracted in ethyl acetate, then washed with Ool

with water, was dried and concentrated to dryness to obtain 220 mg (yield: 36 c/o) acid 7 - (n--tertiary- butoxycarbonyl 6 a-Chloronaphth and 2 a-ylglycylami-spectrum NMR (CDCl ^);

S 1.46 (e, 9:00) jt/2, 15 (two singlets, 3:00) J. 0.3, 35 - ('m, 2:00) 2 - 8, 1 (m-, 8:00).

Having dissolved the product thus formed in 5 ml of trifluoroacetic acid was stirred and the solution at room temperature for 5 min * by evaporation of the solvent and purification of the product by HPLC, was obtained 1 * d-7 - (6 a-chloranapht and 2 a-ylglycylaiflido) 3-methyl 3 a-cephem-to-4-carboxylic,

Example 13

D-7 - (Z naphtylglycylamido ^ O-methoxymethyl s-cephem ^ - carboxylic,

During 5 min, was stirred solution of 570 mg tosylate 7-amino 3 ~méthoxymé-to-polyethylene resins and 3 a-cephem-to-4-carboxylate diphenylmethyl in 30 ml of ethyl acetate containing 10 ml aqueous sodium bicarbonate, then it was concentrated to dryness to obtain the 7"^has in Mino-a 3 a-methoxymethyl-3 a-cephem-to-4-carboxylate diphenylmethyl as a white foam. This foam was dissolved in 20 ml of acetonitrile and the resultant solution was added in one portion to an agitated solution of 301 mg of d n-tert-butoxyearbonyl-a 2 a-naphtylglycine in 20 ml of acetonitrile containing 207 mg of yen, ' n-a-dicyclohexylcarbodiimide and 135 mg of hydroxybenzotriazole. The reaction mixture was stirred during 4 hours at 25 °c * has been poured the mixture into 100 ml of ethyl acetate and the solution was JLavé once with 50 ml aqueous sodium bicarbonate, once with 50 ml of hydrochloric acid In and once with water, then dry and has removed the solvent by evaporation-gous reduced pressure to obtain the D_ 7 - (N-tert butoxybutoxy.carbony 1 and 2 a-naphtylglycylami OD) - 3 a-méthoxyméthy1 and 3 a-cephem-to-4-carboxylate diphenylmethyl as a white foam. This foam was dissolved in 2 ml of triethylsilane and 5 ml of trifluoroacetic acid, the solution was then stirred at 25 °c for 9 min. Removing the solvent by evaporation under reduced pressure, was obtained 26 mg of the trifluoroacetate salt θ of d-7 - (2 a-naphtylglycylamido) - 3 ~méthoxyméthyl-to-~céphem 3 - 4 carboxylic acid,

Having dissolved the salt thus formed in

5 ml of water and 5 ml of acetonitrile and resized the pH of the mixture to 4 * 5 with 1' in-ammonium hydroxide. Was lyophilized solution to obtain a white solid which, after purification by HPLC reverse phase of preparation, gave 20 mg of d-7 - (2 a-naphtylglycylamido) - 3 a-methoxymethyl-3 a-cephem-to-4~carboxylique.

Nuclear magnetic resonance spectrum (dimethyl-sulfoxide d ^): ^ 3, 13 (e, 3:00) J. e 3 * 28 (q-, 2:00) 3/4 * 12 (e, 2:00) 3</4, 95 (D., 1:00) 3/5, 65 (D., 1:00) 3</7, 41 - 4, 23 (m-, 7:00).

Example 14

the n - 7 a-fter -- butoxycarbonyl-(6-methoxy-2-naphthyl) glycylamido] and 3-methyl 3 a-cephem-to-4 carboxylate of P-nitrobenzyl.

Has an agitated solution of 662 mg of n - (2 mm)tertiary- - butoxycarbonyl-(6 a-méthoxy~2-naphthyl) glycine in 100 ml acetonitrile containing 500 mg of d * (2 mm) of n-ethoxycarbonyl 2-ethoxy-L-, 2 a-dihydroquinoline, is added, in one portion, 770 mg (2.2 mm) of 7 ** amino 3 methyl-3 a-cephem-to-4 carboxylate of P-nitrobenzyl. The reaction mixture was stirred at 25 °c for 16 hours, then it was concentrated to dryness to obtain an oil. This oil was dissolved in 50 ml of ethyl acetate and the solution was washed with 25 ml of hydrochloric acid in-, 25 ml aqueous sodium bicarbonate and water * was dried the solution and it was concentrated to dryness to obtain 1.3 g of the n - 7 a-fter -- butoxycarbonyl-(6-methoxy-2-naphthyl) glycylamido] and 3-methyl 3" cephem-to-4 carboxylate of P-nitrobenzyl.

Nuclear magnetic resonance spectrum (CDCl ^);

e 1.39 (e, 9:00) J. (f2_F. 08 and 2.15 (two singlets, 3 Η) I-3.34 (Q-, 2:00) web 3.90 (e, 3 Η) ·, ℮ α, 9 the j (m-, 1:00) 6 5.29 (e, 2:00) web 5.31 (e, 1:00) web 5.68 (m-, 1:00) J. e7, 08 - 8, 25 (the m, 12).

Example 15

D-7" (6-methoxy-2-naphthyl) glycylamido-to-3-methyl 3 - cephem-to-4~carboxylique.

Has a 1.4 g of stirred suspension of palladium-on-carbon 5% in 50 ml of ethanol, was added, in one portion, 1.3 g of 7 a-[n-tert-butoxycarbonyl (6-methoxy-2-naphthyl) glycylamido] and 3-methyl 3 ~céphem and 4 - carboxylate p-nitrobenzyl. The reaction mixture was stirred 3 hours at 25 °c while under a hydrogen pressure of 3.85 kg/cm2. Was then filtered and the reaction mixture was washed the filter cake with fresh ethanol. The filtrate was concentrated to dryness by evaporation under reduced pressure to obtain 1.1 g of acid 7 a-fn-- ter -- butoxycarbonyl-(6-methoxy-2-naphthyl) glycylamido] - 3 a-méthy1 and 3 a-cephem-to-4-carboxylic acid.

Having dissolved acid thus formed in

5 ml of trifluoroacetic acid was stirred and the solution at 25 °c during 5 min. The reaction mixture was added to 20 ml of water and the aqueous solution was lyophilized to obtain 12 hours during the trifluoroacetate of d, L 7 - (6 methoxy 2-naphthylmethyl glycylamido) - 3 methyl-3 a-céphem~4-carboxylic acid. Was redissolved the salt so formed dede.l daps ' fresh water and purified by high performance liquid chromatography to obtain d-7 - (6 methoxy 2 - naphtylglycylamido) - 3 methyl-3 a-cephem-to-4-carboxylic acid.

Nuclear magnetic resonance spectrum (trifluoroacetic acid-to-to dj); (e, 3:00) j * 2, 42; 3.50/(Q-, 2:00) ;

S 4.30 (e, 3:00); S 5.39 (d., 1:00) 5, 8 - 6, 1 (m-, 2:00) Y ((7, 42 - 8, 30 (w, 6:00).

Example 16

By following the general method of examples 14 and 15, which have been reacted - 7^has in Mino-to-3-methyl 3 - cephem-to-4 carboxylate of P-nitrobenzyl with d, L-n -ter -- butoxycarbonyl-(6-hydroxy 2-naphthyl) glycine presence of EEDQ obtain d, L 7 a-n-[ -tertiary- butoxycarbonyl-- (6-hydroxy 2-naphthyl) glycylamido] and 3 methyl-~céphem 3 and 4 carboxylate of P-nitrobenzyl.

Nuclear magnetic resonance spectrum (CDCl ^):

℮ 1.41 (e, 9:00) 5/2, 02 and 2.10 (two singlets, 3:00) j/2,9 - 3.5 (m-, 2:00) 5/4, 9 (m-, 1:00) j/5, 23 (e, 2:00) 3/5, 35 (w, 1:00) 5/5, 6 - 6.0 (m-, 2:00), */ 6, 72 - 8, 21 (the m, 12:00).

Which have been reacted compound thus prepared with hydrogen and palladium-on-carbon 5% to obtain d, L 7 a-[n-tert-butoxycarbonyl (6-hydroxy 2-naphthyl) glycylamido] and 3-methyl 3 a-cephem - 4 - carboxylic acid.

Nuclear magnetic resonance spectrum (CDCl ^):

S 1.15 (e, 9:00) 5 / ΐ, 82 and 1.89 (two singlets, 3 Η) 5 (/ 4, 65 (two doublets, 1:00) j/5, 23 (the m, 1 Η) ; / ό, 0 (m-, 1:00) j/6, 7 - 7, 9 (m-, 7:00) 3/8, 5 (m-, 2:00).

Reacting the compound obtained above with trifluoroacetic acid gave d, L 7 - (6-hydroxy 2 a-naphtylglycylamido) - 3 methyl-3 a-cephem - 4 carboxylic acid. The isomers were separated by high performance liquid chromatography to obtain d-7 - (6-hydroxy 2 a-naphtylglycylamido) - 3 methyl-3 c éphem-a 4 a-CARB SCM oxyli•that

Example 17

D 7 - trifluoroacetate diacid (2 a-naphtylglycylamido) - 3 a-cephem-to-4-carboxylic acid.

A stirred suspension of 2 has a g (10 mm) acid 7"^has in Mino-a 3 a-cephem-a 4 a-cai'cai' boxylic in 15 ml acetonitrile derivative of d *, is added, a portion of the EH, 8 ml (30 mm) of bis - (trimethylsilyl) trifluoroacetamide.

The mixture was stirred 30 minutes at 25 °c to during, and then it was cooled to 0 °c and appended into a portion to an agitated solution of 2 gm (6.6 mm) of d, L-n -ter -butoxycarbonyl 2 a-naphtylglycine in 15 ml D.¹ acetonitrile containing 1.73 grams (7 mm) of EEDQ.

The reaction mixture was stirred for one hour at 25 °c, then it was concentrated to dryness to obtain an oil. This oil was dissolved in 100 ml of ethyl acetate and has been washed four times with portions of 25 ml of hydrochloric acid in vitro, then twice with brine and dry. The solvent is eliminated by evaporation under reduced pressure to give a white foam. This is dissolved in 25 mqusse trirfluoracetic NLF acid has been effected and the solution to a ultrasonic treatment during 5 minutes at 25 °c. The reaction mixture was concentrated to dryness and it was triturated with diethyl ether to obtain 1.8 grams (yield: 55%) of trifluoroacetate of d, L 7 - (2 a-naphtylglycylamido) - 3 a-cephem-to-4-carboxylic acid. It has subjected the resulting product to chromatography on silica gel in c ^ g to reverse phase by eluting with 8 liters of a solution of 1% acetic acid plus a gradient 95% water - 5% /acetonitrile 85 - 15 $$ water acetonitrile (space/space). Suitable fractions have been combined, was then removed the solvent by lyophilization to give 157 mg acid d~7 - (2 a-naphtylglycylamido) - 3 a-cephem-to-4-carboxylic acid.

Absorption spectrum infrared rays (on KBr):

1771>75 cm " * (P-lactam) 3

Nuclear magnetic resonance spectrum (trifluoroacetic acid-to-to dj): GBP 3.55 (q-, 2:00) 4.08 (e, 1:00);

℮ 5, 6 - 6, 0 (m-, 2 Η) ; / 7, 5 - 8, 1 (m-, 7:00).

Example 18

Acid 7 thereof [has-methoxyimino-RO - (8-chloro 2-naphthyl) âcétamido] and 3 a-méthyl~3-to-cephem-to-4-carboxylic acid.

Is transformed 420 mg a-methoxyimino-- (8-chloro 2-naphthyl) acetic acid chloride by reaction with excess chlorine and 500 mg of triphenyl phosphite in 20 ml of dichloromethane. The reaction mixture was added in one portion to a solution of 350 mg of acid stirred 7-amino 3 methyl-3 a-cephem-to-4-carboxylic acid in 5 ml of dichloromethane containing 2 ml of bis (trimethylsilyl) trifluoroacetamide. The reaction mixture was stirred at 25 °c during 6 hours, and then it was diluted by adding 20 ml of methanol " solvent is eliminated by evaporation under reduced pressure to give acid 7 thereof [Œ-to-niéthoxyimino-a-OC - (8-chloro 2-naphthyl) acetamido] and 3-methyl 3 a-cephem - 4 carboxylic acid. Having dissolved the product thus formed in formic acid containing zinc dust metal to obtain, after isolation and purification, acid 7 - (8-chloro 2-naphthyl) glycylamido-to-3-methyl 3 a-cephem-to-4-carboxylic acid.

In the same manner, the following compounds are prepared:

acid 7 - (8-nitrate 2-naphthyl) glycyl-amide 3-chloro 3 a-cephem-to-4~carboxylique 3

acid 7 - (8-hydroxy 2-naphthyl) glycylamido and 3 a-methoxymethyl-3 a-cephem-a 4 a-c arboxyli that,

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

1 * acid 7 "(8-methoxy-2-naphthyl) glycylamido~3" inéthyl and 3 a-cephem-to-4-carboxylic *

The naphtylglycyl cephalosporins obtained by the present invention are antibiotic substances of interest or intermediate products for them. Although active against a broad spectrum of gram-positive and gram-negative, the compounds are particularly effective against a wide variety of gram-positive bacilli. The antibiotic compounds are particularly useful in treating infections caused in animals by the micro gram-positive microorganisms. These compounds are particularly effective for the treatment of upper respiratory tract infections and similar diseases caused by H. influenza vaccine, and S. aureus, S. pyogenes and the like. These compounds are also effective for treating diseases caused by anaerobic cocci such as from Peptostreptococcus anaerobius, Peptostrept. intermedius, Peptostrept. productus, Peptococcussaccharolyticus, w₀prevotii, e. avaerobius, Propionibacterium acnes, Clostridiumand Fusobacteriumor necrophorum analogs.

A preferred specific compound and provided by the present invention is acid 7 - (2 a-naphtylglycylamido) - 3 a-méthy1 and 3 a-cephem-to-4-carboxylic acid (compound of the example 3). The antibacterial activity of this compound and various other compounds of the invention was determined in assays conventional in vitro by diluting the d * agar against a variety of gram-positive microorganisms. The tables below give the minimum inhibitory concentrations specific in a YIG/ml for the compounds when evaluating them against microorganisms indicated. Minimum inhibitory concentrations for different compounds are also known data for comparison purposes.

Table I concentration organism strain

Organism

Reactivity, earworm

Reactivity, methicillin TABLE II

Expanded spectrum - minimum inhibitory concentrations

Organism H * spectrum influ - minimum inhibitory concentrations

Expanded - spectrum concentration

minimum (^ GP/rnl)

Organism strain Cépha - - AFEC

Staphylococcus aureus

A b. fragilis could

Not require reconstitution

Uthe I

TABLE VI

Susceptibility of products anaerobic cocci that isolation of the d * by the method of diluting the d * - agar 24 hours, minimum inhibitory concentrations fyig/ml.) anaerobic micrococcus cefoxitin compound of

1 * 3 g.Peptococcus CCSA -

TABLE VI (result)

The data tables above demonstrate clearly the potent antibacterial activity that exert the compounds provided by the present invention.

In addition to have potent antibacterial activity against a wide variety of microorganisms, in particular, the gram-positive organisms and anaerobic organisms, the 4th compounds the present invention have also demonstrated very favorable pharmacokinetic properties in animals. For example, when acid when administered 7" ^ - ^ - methyl naphtylglycyla-to-midoJ - ^ - ^ - cephem - carboxylic to rats at a dose of 20 mg/kg of intravenous, the concentration in the plasma after one hour was 18.6 ^ GP/ml. ; after 4 hours, it was de 14.1 ^ lg/ml and, after 24 hours, the content in the plasma was always measured at l, 96^pg/ml. This compound is efficiently absorbed orally by rodents and has higher levels of and longer in the blood that cefaclor and cephalexin.

The compounds of the invention also have good stability against P-lactamase inhibitors. The table V gives the results of comparison studies performed on different cephalosporins (the lower digits indicate a higher stability to P-lactamase).

Stability against P-lactamase organism

glycylamido) - 3-me -

the Thyl and 3 a-cephem-a 4 -

carboxylic

Together with their excellent antibacterial activities orally and their excellent stability against of 3 lactamases, favorable pharmacokinetic properties of compounds provided by the present invention in use as agents of particular interest for the treatment of a number of diseases of bacterial origin.

These compounds are particularly well-suited for treatment of patients consulting hospital and, in particular, for subjects with moderate infections of the upper respiratory tract caused by gram-positive microorganisms.

The present invention also provides a T -

means for treating animals suffering from bacterial diseases or suspected of developing a bacterial infection. The method of antibacterial treatment provided may be implemented by administering, to an animal in need of such treatment, an antibiotic of naphtylglycyl-cephalosporin in

an amount effective antibacterial.

This method can be implemented therapeutic or prophylactic benefit, the amount of active antibiotic to be administered will vary depending on the particular compound chosen, the severity of the disease to be treated or prevented, the subject being processed and related factors are commonly encountered in such treatments. However, the compounds will typically be administered in a dose of about 0.5 to about 50 mg/kg of body weight of the animal, more preferably, from about 1 to about 10 mg/kg body weight. These amounts can be administered once or twice a day or more often as needed to treat the disease or the particular subject being processed. A specific daily dose for a human adult means will be between about 200 and about 500 mg/day.

The antibiotic compounds provided by the present invention are active both by oral and parenteral and, accordingly, they may be formulated for any of these modes of administration. Said formulations are another application of the present invention. The formulations of the present invention will comprise about 0.1 to about 95 $by weight of an antibiotic active naphtylglycyl-cephalosporin of the invention admixed with a carrier, diluent or excipient pharmaceutically acceptable carrier therefor. Specific formulations will contain about 10 to about 60 $by weight, more preferably, about 20 to about 50 $by weight of 1' active ingredient.

Convenient for administration orally, the compounds may be mixed with one or the other diluents, excipients and carriers commonly used in formulations for oral administration they can be molded into tablets, dragees or in pills or else they can be coated in gelatin capsules. Among the carriers, diluents and the specific excipients usually used, there are the potato starch, corn starch, sucrose, dextrose, microcrystalline cellulose, dicalcium phosphate, alginic acid and acacia; lubricants such as magnesium stearate J of binding agents such as gum tragacanth or gelatin j of same as flavoring agents such as peppermint oil, flavor cherry or raspberry, wintergreen oil and the like. The compounds may also be formulated as elixirs syrups or d ^ ordinary using diluents such as a fatty oil, methyl - or propyl paraben, flavoring agents and colorants suitable.

The compounds can also be formulated into oral cachet, lozenge or other suitable means to ensure unesidistrifoution sustained and controlled of the active ingredient during a prolonged period.

The antibiotics of the invention may also be formulated for parenteral administration, e.g., intravenous, intramuscular or subcutaneous, as well as transdermally. Such compositions will contain from about 0.1 to about 20 normally% by weight of the active ingredient. Among the excipients, diluents and the specific seats for parenteral formulations, there can be mentioned, for example, an isotonic salt solution, dilute aqueous dextrose, polyhydric aliphatic alcohols or mixtures thereof, for example, glycerin, propylene glycol or polyethylene glycol. Solutions for parenteral administration may also contain preservatives such as phenethyl alcohol, methyl - and propyl paraben or thimerosal. If desired, use can also be about 0.05 to about 0.20# by weight of an antioxidant such as sodium metabisulfite or sodium bisulfite. For use intravenously, in preferred formulations, a preferred initial concentration of from about 0.05 to about 0.25 mg/ml of active ingredient and, for intramuscular injection, a preferred concentration of the active ingredient is between about 0.25 and about 0.50 mg/ml..

The examples below illustrate specific formulations that can be used.

Example 19

The sorbitol solution is added to 40 ml of distilled water and terminates the naphtylglycyl-cephalosporin in suspension. Added and dissolved saccharin, sodium benzoate and the flavoring agent. The volume is adjusted to 100 ml with distilled water. Each ml of syrup contains 5 mg of the naphtylglycyX-cephalosporin antibiotic. This oral formulation is ideally suitable for pediatric use.

Example 20

To 250 mg capsule preparation.

Acid 7~(6" Chloronaphth and 2 a-ylglycyl~amide) - 3 methyl-3 a-cephem-to-4 carboxy -

lic

Of lactose

Corn starch

The ingredients are mixed uniformly

and are coated in gelatin capsules.

These capsules may be orally administered, for example, to the extent of about one or two per day.

Example 21

Preparation of a solution for parenteral administration.

In 700 ml of a solution of propylene glycol and 200 ml of distilled water for injection, is dissolved 20 g of the hydrochloride of d-7 - (2 a-naphtylglycyXamido) - 3 methyl 3 -¹ cephem-to-4-carboxylic acid. Adjusting the pH of the solution to 5j5 with hydrochloric acid and the mixture volume to 1.000 ml with distilled water. The formulation is sterilized and then filled into vials each containing 2 ml 5 ml (either 40 mg) of the active ingredient and the ampules is sealed under a nitrogen atmosphere.

Amount 250 mg

150 mg 100 mg 500 mg