RU2279435C2 - Prodrugs of antibiotic 7-acylamino-3-heteroarylthio-3-cephem carboxylic acid - Google Patents

Abstract

FIELD: organic chemistry, antibiotics, pharmacy.

SUBSTANCE: invention relates to compound of the formula:

wherein R'¹ means hydrogen atom; R'² represents acyl group but with exception of glycyl that is cleaved by mammalian enzyme. Also, invention describes (7R)-7-[(Z)-2-(2-amino-5-chlorothiazol-4-yl)-2-(hydroxyimino)acetamido]-3-{3-[2-N-(L)-aspartylaminoethylthiomethyl]pyrid-4-ylthio}-3-cephem-4-carboxylic acid of the formula:

or its pharmaceutically acceptable salt and pharmaceutical composition used in treatment or prophylaxis of bacterial infection and comprising the effective amount of one or more of indicated compounds. The claimed prodrugs show the enhanced solubility in water at neutral or near to neutral pH value as compared with the parent compounds.

EFFECT: improved and valuable properties of prodrugs.

15 cl

Description

FIELD OF THE INVENTION

The present invention relates to organic chemistry, medical chemistry, biochemistry and medicine. In particular, it relates to prodrugs of cephalosporin antibiotics, when said prodrugs are significantly more soluble in water than the parent antibiotic, which, among other advantages, has the advantage of parenteral administration. Prodrugs are readily converted under physiological conditions to give the starting antibiotic.

State of the art

The following description provides an understanding of the present invention. Over the past three decades, many antibiotics have become available for clinical use. However, the mixed use of these antibiotics has increased the resistance of a number of clinically important bacterial strains such as Salmonella, S. pneumoniae, Enterobacteriaceae, Staphylococcus aureus and Pseudomonas. The rate of sustainability has increased dramatically in recent years. Among the diverse classes of antibiotics that are characterized by an increase in resistance growth, cephalosporins were particularly prominent.

Immediately after 1965, more than 70 cephalosporins became available for clinical use in the treatment of bacterial infections. Their mode of action involves the inhibition of bacterial peptidoglycan biosynthesis, which is an important step in creating a cell membrane. Resistance to cephalosporins is determined primarily by three mechanisms: (a) through the development of β-lactamases, which split β-lactam rings, thus deactivating the antibiotic; (b) through changes in the composition of the cell wall, which lead to a decrease in the penetration of the antibiotic; and (c) through changes in penicillin-bound proteins (PBPs), leading to a weakening of the binding of PBPs to β-lactams, where binding is essential for inhibiting cell membrane biosynthesis with β-lactams.

Examples of bacteria that are resistant due to insufficient PBP binding are methicillin-resistant Staphylococcus aureus ("MRSA") and Enterococci. Resistance in MRSA is due to the presence of high levels of altered PBP, PBP2a, which show low binding to β-lactams. Attempts to circumvent this resistance mechanism have led to the discovery of many new antibacterial compounds.

Compounds that are shown to be effective against bacteria whose resistance is determined by low binding to PBP include those described in pending International Application PCT / US95 / 03976 and in patent applications US 08/222262, April 1 1994; 08/369798, dated January 6, 1995; 08/413713, 08/413714, 08/415065, 08/413712, 08/415064 and 08/415069, all from March 29. 1995; 08/455969, dated May 31, 1995; 08/457673, dated June 1, 1995; 08/940508 and 08/937812, both of September 29, 1997; 08/730041, 08/730039, 08/728232, 08/430042, 08/728233 and 08/730040, all of October 11, 1996 and 08/842915, of April 17, 1997; all of them are included in the present description by reference, including all figures.

A class of compounds that has demonstrated activity against β-lactams, especially cephalosporin-resistant bacteria, relates to the 7-acylamino-3-heteroarylthio-3-cefemcarboxylic acids described in US 6,025,352, which is also incorporated by reference in the present description, including any drawings. Among the 7-acylamino-3-heteroarylthio-3-cefemcarboxylic acids, the most active compound is (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido ] -3- {3- (2-aminoethylthiomethylpyrid-4-ylthio) -3-cephem-4-carboxylic acid (I):

However, demonstrating a high level of activity against β-lactam-resistant bacterial strains in vitro, the use of 1 in vivo is limited due to its relatively low solubility in water, which in turn limits the available methods of administration. That is, the solubility of 1 in water at or near a neutral pH value, for example a pH of from about 5 to about 7, is insufficient.

There is a need for derivative 1, which is better soluble in water at a pH near or neutral, but which has the same high bactericidal activity as 1, or which easily transfers to compound 1 itself in vivo. The present invention provides such a derivative 1.

SUMMARY OF THE INVENTION

The present invention relates to prodrugs 1 that exhibit surprising solubility in water and are readily bio-convertible in vivo to a precursor compound.

Thus, in one aspect, the present invention relates to compounds of the formula:

Where:

R ^{′ 1 is} selected from the group consisting of hydrogen and —C (O) CH (NH ₂ ) CH ₃ ; and

R ^{′ 2 is} selected from the group consisting of hydrogen and an acyl group that is cleaved by an enzyme found in mammals.

In another aspect, the present invention relates to the described compounds, where:

R ^{′ 2 is} selected from the group consisting of hydrogen, —C (O) —R ⁸⁸ , —C (O) —OR ⁸⁹ , —C (O) —CH (NHR ′ ³ ) —alk ₄ , and

Where:

или

R ⁸⁸ represents

or

R ^{′ 3 is} selected from the group consisting of hydrogen, —C (O) —OR ⁸⁹ and —C (O) —CH (NH ₂ ) —alk ₄ ;

; гдеalk _{4 is} selected from the group consisting of hydrogen and optionally substituted alkyl, wherein said alkyl is optionally substituted with one or more substituents selected from the group consisting of hydrogen, phenyl, —COOH, —C (O) —OR ⁸⁹ , —C ( O) NH ₂ -, -OH, -SH, -NH ₂ , and

; Where

R ^{89 is} selected from the group consisting of benzhydryl, tert-butyl, allyl, p-nitrobenzyl, benzyl, n- or o-nitrobenzyl, 2,2,2-trichloroethyl, cinnamyl, 2-chloroallyl, tert-amyl, trityl, 4 -methoxytrityl, 4,4'-dimethoxytrityl, trimethylsilyl, tert-butyldimethylsilyl, β- (trimethylsilyl) ethyl, 4- or 2-methoxybenzyl, 2,4-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, methoxymethyl and 3,3-dimethylallyl.

In another aspect, the present invention relates to the compounds described above, in which R ^{′ 3 is} selected from the group consisting of hydrogen, methyl and —C (O) CH (NH ₂ ) CH ₃ .

.In a further aspect, the present invention relates to the compounds described above in which alk _{4 is} selected from the group consisting of hydrogen, —CH ₃ , —CH (CH ₃ ) ₂ , —CH ₂ OH, —CH ₂ NH ₂ , —CH ₂ CH ₂ NH ₂ , -CH ₂ CH ₂ CH ₂ NH ₂ , -CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ , -CH ₂ COOH, -CH ₂ CH ₂ COOH, -CH ₂ -C (O) NH ₂ , - CH ₂ CH ₂ —C (O) NH ₂ , and

.

Pharmaceutically acceptable salts of any of the compounds described above are also an object of the present invention.

An object of the present invention is also a compound selected from the group consisting of:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ornithylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -prolylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-N- (L) -ananylamino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-aminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L, L) -alanylalanaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N-glycylaminoethylthiomethyl] pyrid-4 -ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -aspartylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ananylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) - ( N _α- methyl) alanylaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -histidylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -valaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -aspartylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -lisylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -serylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -glutamylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -glutaminaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N - ((5-methyl -1,3-dioxolan-4-en-2-one-4-yl) methoxycarbonyl) aminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid:

Another object of the present invention is a compound selected from the group consisting of:

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ornithylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid,

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L, L) -alanylalanaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid,

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -aspartylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid,

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ananylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid,

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -lisylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid,

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -glutamylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cefem-4-carboxylic acid; and,

(7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N - ((5-methyl -1,3-dioxolan-4-en-2-one-4-yl) methoxycarbonyl) aminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid.

Finally, an object of the present invention is the compound (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [ 2-N- (L) -aspartylaminoethylthiomethyl] pyrid-4-ylthio} -3-cephem-4-carboxylic acid:

In addition, the present invention relates to a pharmaceutically acceptable salt of any of the compounds of the present invention.

Another object of the present invention is a composition for use in the treatment of bacterial infection in a patient, comprising one or more compounds of the present invention.

Another object of the present invention is a composition comprising a compound of the present invention and a pharmaceutically acceptable carrier or excipient.

The object of the present invention is also a composition for use in the treatment of bacterial infections in humans, comprising one or more compounds of the present invention.

The present invention also relates to a composition for use in the treatment of bacterial infection in humans, further comprising a pharmaceutically acceptable carrier or excipient.

In one aspect, the present invention relates to the compositions described above, which are useful in the treatment of bacterial infections resistant to β-lactam antibiotics.

Β-lactam antibiotic-resistant bacterial infections are bacterial infections that are resistant to methicillin, ampicillin or vancomycin, which is an aspect of the present invention.

In addition, those bacterial infections caused by Staphylococcus or Enterococcus species of bacteria are another aspect of the present invention.

Staphylococcus or Enterococcus species are resistant to some or all other β-lactam antibiotics in another aspect of the present invention.

Resistant Staphylococcus species are S.aureus Col (Meth ^R ) (bla-), S.aureus 76 (Meth ^R ) (bla +), S.aureus ATCC 29213, S.Aureus ATCC 25913, S.Aureus ATCC 32432 or S. Aureus CoIBA in a further aspect of the present invention.

Resistant Enterococcus species are E. faecium ATCC 35667 or E. facialis ATCC 29212 in one aspect of the present invention.

And finally, compositions intended for use in the prophylaxis of patients to prevent bacterial infections are also an aspect of the present invention.

DETAILED DESCRIPTION OF THE INVENTION

Brief Description of Drawings

The drawing shows the water solubility of the original antibiotic, compound 1 in its zwitterionic form (2) in comparison with its prodrug of the present invention - compound 3:

Definitions

As used in the present invention, the term "alkyl" refers to saturated aliphatic hydrocarbon, straight and branched groups. Preferably, the alkyl group has 1 to 20 carbon atoms (in the case where the numerical range is indicated, for example “1-20”, this means that the group, in this case the alkyl group, may contain 1 carbon atom, 2 carbon atoms , 3 carbon atoms and so on, up to and including 20 carbon atoms). More preferably, the average size of the alkyl is from 1 to 10 carbon atoms. More preferably, alkyl is lower alkyl having from 1 to 4 carbon atoms. The alkyl group may be substituted or unsubstituted. Examples, without limitation, unsubstituted alkyl groups are methyl, ethyl, n-propyl, iso-propyl, n-butyl, sec-butyl, iso-butyl, tert-butyl and 2-methylpentyl. In the case of substitution, the substituent group (s) may be, without limitation, one or more independently selected from the group consisting of halogen, hydroxy, cyano, nitro, lower alkoxy, carboxy, carbonyl, alkylcarbonyl, alkoxycarbonyl, mercapto, alkylthio, amino, amido, isothioureido, amidino, guanidino, optionally substituted aryl, optionally substituted aryloxy, optionally substituted arylthio, optionally substituted 6-membered heteroaryl having from 1 to 3 nitrogen atoms in the ring, optionally substituted 5-membered hetero aryl having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur and optionally substituted by a 5- or 6-membered heteroalicyclic group having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen oxygen and sulfur. Examples of, but not limited to, substituted alkyl groups include trifluoromethyl, 3-hydroxyhexyl, 2-carboxypropyl, 2-fluoroethyl, carboxymethyl, 4-cyanobutyl, 2-guanidinoethyl and 3-N, 'N'-dimethylisothiouronium propyl.

A “cycloalkyl” group refers to a 3-8 membered all-carbon monocyclic ring and an all-carbon 5-membered / 6-membered or 6-membered / 6-membered fused bicyclic ring or a multicyclic fused ring (“fused” ring system means that each the ring in the system has an adjacent pair of carbon atoms with each other ring in the system) group, where one or more rings may contain one or more double bonds, but none of the rings has a completely conjugate π-electron system. Examples, without limitation, of cycloalkyl groups are cyclopropane, cyclobutane, cyclopentane, cyclopentene, cyclohexane, cyclohexadiene, adamantane, cycloheptane and cycloheptatriene. The cycloalkyl group may be substituted or unsubstituted. In the case of substitution, the substituent (s) group (s) may be, without limitation, one or more independently selected from the group consisting of halogen, hydroxy, alkoxy, mercapto, alkylthio, cyano, nitro, optionally substituted aryl, optionally substituted aryloxy, mercapto , alkylthio, optionally substituted arylthio, amino, amido, carboxy, carbonyl, alkylcarbonyl, alkoxycarbonyl, optionally substituted 6-membered heteroaryl having from 1 to 3 nitrogen atoms in the ring, optionally substituted 5-membered heteroaryl, having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur and an optionally substituted 5- or 6-membered heteroalicyclic group having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur.

An "alkenyl" group refers to an alkyl group, as defined in the present invention, consisting of at least two carbon atoms and at least one carbon-carbon double bond.

An "alkynyl" group refers to an alkyl group, as defined herein, consisting of at least two carbon atoms and at least one carbon-carbon triple bond.

An “aryl” group refers to a fully carbon monocyclic or fused polycyclic ring (that is, the ring of which has a common pair of carbon atoms) group having a fully conjugated π-electron system. Examples, without limitation, of aryl groups are phenyl, naphthalenyl and anthracenyl. The aryl group may be substituted or unsubstituted. In case of substitution, the substituent (s) group (s) may be without limitation one or more independently selected from the group consisting of alkyl, (halogen) ₃ C-, halogen, hydroxy, alkoxy, mercapto, alkylthio, cyano, nitro, amino, amido, carboxy, carbonyl, alkylcarbonyl, alkoxycarbonyl, optionally substituted 6-membered heteroaryl having 1 to 3 nitrogen atoms in the ring, optionally substituted 5-membered heteroaryl having 1 to 3 heteroaryls in the ring selected from the group consisting of nitrogen oxygen and sulfur, and optionally substituted a 5- or 6-membered heteroalicyclic group having from 1 to 3 heteroatoms in the ring selected from the group consisting of nitrogen, oxygen and sulfur. Examples, without limitation, of substituted aryl groups are biphenyl, yodobifenil, methoxybiphenyl, anthryl, bromophenyl, iodophenyl, chlorophenyl, hydroxyphenyl, methoxyphenyl, formylphenyl, acetylphenyl, trifluoromethylthiophenyl, trifluoromethoxyphenyl, alkylthiophenyl, trialkilammoniyfenil, amidofenil, tiazolilfenil, oksazolilfenil, Imidazolylphenyl, imidazolilmetilfenil, cyanophenyl, pyridylphenyl, pyrrolylphenyl, pyrazolylphenyl, triazolylphenyl and tetrazolylphenyl.

As used in the present invention, a “heteroaryl” group refers to a monocyclic or fused ring (i.e., rings have an adjacent pair of atoms) group having in the ring (s) one or more atoms selected from the group consisting of nitrogen, oxygen and sulfur, and additionally having a conjugated π-electronic system. Examples, without limitation, of heteroaryl groups are pyrrole, furan, thiophene, imidazole, oxazole, thiazole, pyrazole, pyridine, pyrimidine, quinoline, isoquinoline, purine and carbazole. A heteroaryl group may be substituted or unsubstituted. In case of substitution, the substituent (s) group (s) may be, without limitation, one or more independently selected from the group consisting of alkyl, (halogen) ₃ C-, halogen, hydroxy, lower alkoxy, mercapto, alkylthio, arylthio, cyano, nitro, amino, amido, carboxy, carbonyl, alkylcarbonyl and alkoxycarbonyl. Examples of substituted heteroaryl groups are, without limitation, 2-aminothiazol-4-yl, 2-amino-5-chlorothiazol-4-yl, 5-amino-1,2,4-thiadiazol-3-yl, 2,3-dioxopiperazinyl , 4-alkylpiperazinyl, 2-iodo-dibenzfuranyl and 3-hydroxy-4-dibenzthienyl.

A "heteroalicyclic" group refers to a monocyclic or fused ring group having in the ring (s) one or more atoms selected from the group consisting of nitrogen, oxygen and sulfur. Rings may also have one or more double bonds. However, rings do not have a fully conjugated π-electron system. The heteroalicyclic ring may be substituted or unsubstituted. In case of substitution, the substituent (s) group (s) may be, without limitation, one or more independently selected from the group consisting of alkyl, (halogen) ₃ C-, halogen, hydroxy, alkoxy, mercapto, alkylthio, cyano, nitro, amino , amido, carboxy, carbonyl, alkylcarbonyl and alkoxycarbonyl.

The “Hydroxy” group refers to the —OH group.

An “alkoxy” group refers to both an —O— (unsubstituted alkyl) and an —O— (unsubstituted cycloalkyl) group.

An “aryloxy” group refers to both an —O-aryl and an —O-heteroaryl group as defined herein.

The “mercapto” group refers to the —SH group.

The "alkylthio" group refers to both the (unsubstituted alkyl) S- and the (unsubstituted cycloalkyl) S-group.

The "arylthio" group refers to both -S (aryl) and -S (heteroaryl group), as defined in the present invention.

"Halogen" refers to fluorine, chlorine, bromine or iodine.

The “cyano” group refers to the —C≡N group.

The “nitro” group refers to the —NO ₂ group.

An "amino" group refers to an NRR 'group, where R and R' are independently selected from the group consisting of hydrogen, alkyl, cycloalkyl, aryl, heteroaryl and a heteroalicyclic group.

The "amido" group refers to —C (O) NRR ′, where R and R ′ are as defined above.

A “carboxy” group refers to a —CO (O) H group.

A “carbonyl" group refers to a —C (O) H group.

An “acyl” group refers to a —C (O) R group in which R is hydrogen or alkyl, as defined above, such as formyl, acetyl, propionyl or butyryl.

An “alkylcarbonyl” group refers to a —C (O) (alkyl) group.

An “alkoxycarbonyl” group refers to a —C (O) (O-alkyl) group.

“Trialkylsilyl” groups refer to the RR′R ″ Si group, where R, R ¹ and R ″ are alkyl as defined above.

The "trialkylammonium" group refers to [PR'R''N-] ⁺ , where R, R 'and R "are alkyl, as defined above.

The term "method" refers to methods that disclose techniques and procedures for performing the task, including, but not limited to those methods, techniques and procedures that are known to every specialist or can easily be developed from the specified methods, techniques and procedures, well-known secondary specialists in the chemical, pharmaceutical, biological, biochemical and medical fields of technology.

"Pharmaceutical composition" means a mixture of one or more of the compounds described in the present invention, or its physiologically acceptable salts, with other chemical components, such as physiologically acceptable carriers and excipients. The aim of the pharmaceutical composition is to facilitate the administration of a compound to the body.

As used in the present invention, a pharmaceutically acceptable salt refers to any of the compounds of the present invention in its cationic or anionic form together with the corresponding counterion. The compound of the present invention may exist as a separate or doubly charged particle, in the latter case it will form a salt with two counterions, which may be the same or different. Preferred pharmaceutically acceptable salts include (1) inorganic salts such as sodium salt, potassium salt, chloride, bromide, iodide, nitrate, phosphate or sulfate; (2) carboxylic acid salts such as acetate, trifluoroacetate, propionate, butyrate, maleate or fumarate; (3) alkyl sulfonates such as methanesulfonate, ethanesulfonate, 2-hydroxyethyl sulfonate, n-propyl sulfonate or isopropyl sulfonate; and (4) hydroxycarboxylates such as lactate, malate and citrate. Salts in which the compound of the present invention is in the form of a cation are prepared by reacting the compound with an organic or inorganic acid, such as hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid, phosphoric acid, methanesulfonic acid, ethanesulfonic acid, p-toluenesulfonic acid, salicylic acid and the like. The acid used provides the anion. Salts in which the compound of the present invention is an anion are prepared by reacting any compound of the invention with an organic or inorganic base such as benzatene, chloroprocaine, choline, diethanolamine, ethylenediamine, meglumine, procaine and hydroxide, alkoxide, carbonate, bicarbonate, sulfate, bisulfate , amide, alkylamide or dialkylamide salts of lithium, sodium, potassium, magnesium, calcium, aluminum and zinc. The base provides a cation.

“Prodrug” refers to a compound that is converted to an in vivo precursor drug. Prodrugs are often useful in some situations, as they can be administered more easily than the parent drug. They may, for example, be bioavailable for oral administration, while the parent drug is not. A prodrug may also have improved solubility in pharmaceutical compositions as compared to precursor drugs. An example of a prodrug, without limitation, may be a compound of the present invention that is administered as an ester (“prodrug”) to facilitate penetration through the cell membrane, where solubility in water is undesirable in terms of mobility, but then metabolically hydrolyzes to a carboxylic acid, active principle , already inside the cell, where solubility in water is an advantage.

Another example of a prodrug can be a single amino acid or a short polypeptide, for example, without limitation, a polypeptide of 2-10 amino acids linked through a terminal amino group to a carboxyl group of a compound or via a carboxyl group of an amino acid to an amino group of a compound, as is the case with the compounds of the present invention . The compound thus formed is then converted in vivo to isolate the active molecule.

As used in the present invention, a “physiologically acceptable carrier” refers to a carrier or diluent that does not cause significant irritation in the body and does not reduce the biological activity and properties of the administered compound.

"Excipient" refers to an inert substance that is added to the pharmaceutical composition, which further facilitates the administration of the compound. Examples of excipients, without limitation, include calcium carbonate, calcium phosphate, a variety of sugars and types of starch, cellulose derivatives, gelatin, vegetable oils and polyethylene glycols.

"In vitro" refers to processes carried out in an artificial environment such as, for example, without limitation, a test tube or Petri dish containing a culture medium.

"In vivo" refers to processes occurring in a living organism, such as, without limitation, a mouse, rat, dog, cat, rabbit, cow, pig, horse or human.

As used in the present invention, “bacterial infection” refers to the formation of a significant population of pathogenic bacteria in a patient that adversely affects the health and well-being of the patient and / or leads to the appearance of visible symptoms associated with certain bacteria.

As used in the present invention, the terms “prevent”, “prevention” and “prophylaxis” refer primarily to a method of protecting an organism from bacterial infections.

As used in the present invention, the terms “treat”, “treatment” and “treatment” refer to a method of alleviating or stopping the effects of bacterial infections and / or their associated symptoms.

As used in the present invention, “administer”, “administration” or “administration” refers to the delivery of a compound or its salt or pharmaceutical composition containing the compound or its salt to the body in order to prevent or treat bacterial infections.

The term "patient" refers to any living organism that can be infected with bacteria. In particular, “patient” refers to mammals, such as a dog, cat, horse, cow, pig, rabbit, goat or sheep. In particular, the term "patient" in the present invention refers to a person.

The term “therapeutically effective amount”, as used in the present invention, refers to that amount of the administered compound that alleviates to some extent one or more of the symptoms found in the treatment. Regarding the treatment of bacterial infections, a therapeutically effective amount refers to that amount of a compound that (1) reduces, preferably stops the growth of bacteria in the patient’s body, (2) inhibits (i.e. slows down, preferably, stops) the growth of bacteria, (3 ) inhibits to some extent (i.e., slows to some extent, preferably stops) the spread of infection caused by bacteria and / or, (4) reduces to some extent (preferably, strange) one or more symptoms associated with a bacterial infection.

The term "prophylactically effective amount" refers to that amount of a compound that (1) maintains a reduced level of the bacterial population; (2) maintains a level of inhibition of bacterial reproduction; (3) maintains a level of inhibition of the spread of infection; and / or (4) maintains a level of alleviation of one or more symptoms associated with a bacterial infection achieved by administering a therapeutically effective amount of a compound.

The term "beta-lactam resistant bacteria" refers to bacteria against which the beta-lactam antibiotic has a minimum inhibitory concentration (MIC) of greater than 32 mg / ml.

Synthesis

Compound 1 is synthesized using reactions and procedures from readily available materials that are well known in organic chemistry. Descriptions such as (without limitation) March, Advanced Organic Chemistry: Reactions, Mechanisms and Structure (McGraw-Hill, latest edition); Larock, Comprehensive Organic Transformations (VCH Publishers, latest edition); Greene and Wuts, Protective Groups in Organic Synthesis (John Wiley & Sons, latest edition) and GIGeorg, The Organic Chemistry of β-Lactams (VCH Publishers, latest edition), reveal quite fully the direction of synthesis 1. Production method 1 may also be found in US 6025352, which is fully incorporated by reference in the present description, including any drawings. The following examples are typical for such a synthesis:

A. (Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetic acid.

To a solution of (Z) -2- (2-aminothiazol-4-yl) -2- (triphenylmethoxyimino) acetic acid (5.81 g, 13.47 mmol) in DMF (30 ml), N-chlorosuccinimide (1.80 g, 13.47 mmol). After stirring overnight, the reaction mixture was poured into water (500 ml), and the resulting precipitate was filtered off, washed with water and ethyl acetate and dried in vacuo to obtain 4.43 g (71%) of the title compound: ¹³ C NMR (CDCl ₃ ) δ 108.5, 125.6, 126.2, 126.6, 127.3, 134.7, 141.8, 146.5, 162.1, 163.3.

B. Diphenylmethyl ester (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3-chloro-3-cefem-4- carboxylic acid.

To a solution of the salt of toluenesulfonic acid diphenylmethyl ester of 7-amino-3-chlorocephalosporic acid (5.0 g, 8.72 mmol) in dry THF (100 ml) at room temperature, pyridine (0.63 g, 10.0 mmol) is added, then (Z) -2- ( 2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetic acid (5.81 g, 13.47 mmol). The resulting suspension was cooled to -15 ° C and pyridine (1.42 g, 22.5 mmol) was added additionally, then phosphorus oxychloride (1.64 g, 17.5 mmol) was added dropwise, keeping the reaction temperature below -10 ° C. After 30 minutes, ethyl acetate (200 ml) was added, followed by water (150 ml). The aqueous layer was carefully extracted with ethyl acetate and the combined organic extracts were dried over sodium sulfate and concentrated in vacuo to give a crude product which was purified by flash chromatography on a silica gel column (ethyl acetate / hexane 3: 1) to obtain the title compound (5.37 g, 65%): ¹ H NMR (CDCl ₃ / CD ₃ OD) δ 3.35 (d, 1H, J = 18), 3.68 (d, 1H, J = 18), 5.07 (d, 1H, J = 5), 5.80 (broad singlet, 2H), 6.04 (dd, 1H, J = 9.5), 7.03 (s, 1H), 7.06 (d, 1H, J = 9), 7.22-7-50 (m, 25H).

C. Diphenylmethyl ester (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3-mercapto-3-cefem-4- carboxylic acid.

To a solution of diphenylmethyl ether (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3-chloro-3-cefem-4 -carboxylic acid (4.0 g, 4.72 mmol) in DMF (30 ml) at -20 ° C, powdered sodium hydrosulfide hydrate (1.10 g, 19.6 mmol) is added in one go. After 15 minutes, the reaction mixture was poured into a 0.5 M sodium monophosphate solution (approximately 100 ml). The mixture was extracted with ethyl acetate and the organic layer was washed thoroughly with water. After concentration in vacuo, the crude title product is obtained in the form of a yellow foam 3.8 g (95%):

¹ H NMR (CDCl ₃ / CD ₃ OD) δ 3.38 (d, 1H, J = 15), 4.43 (d, 1H, J = 15), 5.03 (d, 1H, J = 5), 5.80 (d, 1H , J = 5), 5.99 (broad singlet, 1H), 6.80 (s, 1H), 7.05-7.50 (m, 25H).

D. Hydrochloride-chloromethyl-4-chloropyridine

Thionyl chloride (0.714 ml, 9.78 mmol) was added at room temperature in dry DMF (7 ml). After 30 minutes, the solution was transferred through a cannula into a solution of 3-hydroxymethyl-4-chloropyridine (700 mg, 4.89 mmol) in DMF (3 ml). After 45 minutes, the product was precipitated by the addition of dry ether (100 ml), washed with ether and then dried in vacuo to give 813 mg (84%) of the title compound:

¹ H NMR (CD ₃ OD) δ 5.00 (s, 2H), 8.31 (d, 1H, J = 5), 8.99 (d, 1H, J = 5), 9.18 (s, 1H).

E. 3- (N-tert-Butoxycarbonylaminoethylthiomethyl) -4-chloropyridine.

To a solution of 3-chloromethyl-4-chloropyridine hydrochloride (513 mg, 2.59 mmol) in DMF (6 ml) was added sodium iodide (386 mg, 2.59 mmol), diisopropylethylamine (1.12 ml, 6.47 mmol) and 2- (N tert-butoxycarbonylamino) ethanethiol (458 mg, 2.59 mmol). After 2 hours, the reaction mixture was partitioned between diluted HCl and ethyl acetate. The organic layer is washed with water, dried over sodium sulfate and concentrated, which gives 750 mg of an oily product (96%), which is used in the next step without further purification:

¹ H NMR (CDCl ₃ ) δ 1.43 (s, 9H), 2.61 (m, 2H), 3.35 (m, 2H), 3.81 (s, 2H), 490 (broad singlet, 1H), 7.35 (d, 1H, J = 4), 8.40 (d, 1H, J = 4), 8.57 (s, 1H).

F. Diphenylmethyl ester (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3- [3-N-tert-butoxycarbonylaminoethylthiomethyl ) pyrid-4-ylthio] -3-cefem-4-carboxylic acid.

To a solution of diphenylmethyl ether (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3-mercapto-3-cefem-4-carbon acids (650 mg, 0.777 mmol) in DMF (3 ml) add 3- (N-tert-butoxycarbonylaminoethylthiomethyl) -4-chloropyridine (242 mg, 0.80 mmol) at room temperature. After stirring overnight, the reaction mixture was partitioned between water and ethyl acetate. The organic layer was washed thoroughly with water, dried over sodium sulfate and concentrated to give a crude product, which was purified by silica gel radial chromatography (dichloromethane / methanol; v / v, 50/1) to obtain 220 mg of the title compound (26% ):

¹ H NMR (CDCl ₃ / CD ₃ OD) δ 1.23 (s, 9H), 2.32 (t, 2H, J = 6), 2.98 (d, 1H, J = 18), 3.06 (m, 2H), 3.40 ( d, 1H, J = 18), 3.46 (s, 2H), 5.03 (d, 1H, J = 5), 5.52 (broad singlet, 1H), 5.94 (d, 1H, J = 5), 6.80 (s, 1H), 6.90 (d, 1H, J = 6), 7.00-7.22 (m, 25H), 8.01 (d, 1H, J = 6), 8.08 (s, 1H).

G. Methanesulfonic acid salt (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- [3- (aminoethylthiomethyl) pyrid- 4-ylthio] -3-cefem-4-carboxylic acid.

To a cooled solution (0 ° C) of diphenylmethyl ether (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3- [3- (N-tert-butoxycarbonylaminoethylthiomethyl) pyrid-4-ylthio] -3-cephem-4-carboxylic acid (1.00 g, 0.907 mmol) in dichloromethane (10 ml) and anisole (1.0 ml) add trifluoroacetic acid (13 ml). After 1.5 hours, the reaction mixture was concentrated in vacuo at room temperature and the oily residue was dissolved in 98% formic acid (20 ml). After 4 hours at room temperature, formic acid was removed in vacuo and the residue was dissolved in water (25 ml). Insoluble product is removed by centrifugation. The supernatant was purified on an HP20 column, eluting with water, then with 0.1 M aqueous ammonium acetate, and then, finally, the product was eluted with 1: 4 acetonitrile / water. The eluate was concentrated to about 1/10 of the original volume and the resulting precipitate was filtered off, washed with water and dried in vacuo to give a zwitterionic product (260 mg). The methanesulfonate salt is obtained by suspending the zwitterionic product in water (15 ml), followed by the addition of methanesulfonic acid (1.0 M in water, 0.98 equiv.) And acetonitrile (5 ml). After evaporation, the resulting solution was dried, the residue was dissolved in water (30 ml) and centrifuged to remove the insoluble product, and the supernatant was lyophilized to give the title compound (274 mg, 44%).

¹ H NMR (D ₂ O) δ 3.11 (s, H), 3.19 (m, 2H), 3.52 (m, 2H), 3.67 (d, 1H, J = 17), 4.22 (d, 1H, J = 17), 4.33 (s, 2H), 5.76 (d, 1H, J = 4), 6.29 (d, 1H, J = 4), 7.93 (d, 1H, J = 4), 8.78 (d, 1H, J = 4), 8.87 (s, 1H).

The prodrugs 1, which are the subject of the present invention, can also be obtained using the standard methods of organic synthesis described in the above articles. The following are examples of such preparation. The demonstrated synthesis should not be construed as limiting the scope of the present invention in one way, whatever it may be. That is, other prodrugs and other approaches to their synthesis will become apparent to those skilled in the art based on the present invention; these prodrugs and their synthesis are within the scope of the present invention.

1. The double salt of (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- ( L) -α-glutamylaminoethylthiomethyl] pyrid-4-ylthio} -3-cephem-4-carboxylic acid with trifluoroacetic acid

A solution of diphenylmethyl ester (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (triphenylmethoxyimino) acetamido] -3-mercapto-3-cefem-4-carboxylic acid (354 mg, 0.41 mmol) and 4-chloro-3- {2- [N- (L) -α- (N-tert-butoxycarbonyl) - (γ-tert-butoxy) glutamyl] aminoethylthiomethyl} pyridine (183 mg , 0.41 mmol) in DMF (2.5 ml) was stirred under nitrogen for 3 hours and diluted with ethyl acetate (25 ml). The reaction mixture was washed thoroughly with water and dried over anhydrous sodium sulfate. The mixture was passed through a silica gel chromatography column (eluting with hexane / ethyl acetate at 2: 1, then at 1: 3) to obtain the desired condensation product as an oily solid (107 mg). Deprotection is carried out under standard conditions (trifluoroacetic acid, dichloromethane, triethylsilane) and the title product is precipitated by the addition of diethyl ether. The precipitate was filtered off, washed thoroughly with diethyl ether and dried to obtain the title compound (48 mg).

¹ H NMR (CD ₃ OD): δ 2.25-2.37 (m, 2H), 2.65 (t, 2H, J = 8), 2.80-2.96 (m, 2H), 3.58-3.78 (m, H), 4.08- 4.20 (m, 4H), 5.58 (d, 1H, J = 8), 6.20 (d, 1H, J = 8), 7.80 (d, 1H, J = 8), 8.62 (d, 1H, J = 8) 8.91 (s, 1H).

2. (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -α- (N-tert-butokeicarbonyl) - (γ-tert-butoxy) asparagylaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid

To a stirred suspension of (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-aminoethylthiomethyl] pyrid- 4-ylthio} -3-cefem-4-carboxylic acid (8.6 g, 14.28 mmol) in DMF (25 ml), triethylamine (2.0 ml, 14.5 mmol) was added at room temperature, and then a solution of N-tert-butoxycarbonyl- [ α- (N-hydroxysuccinimide) ester] -γ-tert-butyl ester of aspartic acid (5.5 g, 14.28 mmol) in DMF (25 ml). After 30 minutes, ice water (200 ml) was added to the reaction mixture, followed by hydrochloric acid (1.0 M, 14.3 ml). A precipitate formed, which was filtered off, washed thoroughly with water and dried under reduced pressure to give the crude product as a yellowish solid (10.0 g).

3. The sodium salt of (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- ( L) -γ-asparagylaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid.

To a stirred suspension of crude (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- ( L) -α- (N-tert-butoxycarbonyl) - (γ-tert-butoxy) -asparagylaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid (11.8 g, 13.52 mmol) in dichloromethane (200 ml ) triethylsilane (66 ml) is added at room temperature, followed by trifluoroacetic acid (200 ml). The resulting clear solution was stirred for 2.5 hours. After removing the solvents under reduced pressure, the oily residue was redissolved in ethyl acetate (50 ml) and the trifluoroacetate salt of the product was precipitated by adding diisopropyl ether (100 ml). The precipitate was filtered off, washed thoroughly with diisopropyl ether and dried under reduced pressure to obtain the crude salt of trifluoroacetate as a yellow solid (15.0 g). A solution of the crude product (in 600 ml of water was adjusted to pH 6.0 with an aqueous solution of sodium bicarbonate) was loaded onto a reverse phase preparative column, which was eluted with an acetonitrile / phosphate buffer mixture. The fractions containing the pure product (800 ml) were combined, the acetonitrile was removed by evaporation under reduced pressure, and the aqueous solution of the product was freed from the salt by loading onto a column containing HP20 resin and thoroughly washed with water. The product was eluted with 20% aqueous acetonitrile and the fractions containing the pure, salt-free product were concentrated under reduced pressure to remove acetonitrile. The remaining water is removed from the resulting turbid solution by lyophilization. The lyophilized product is redissolved in water and the pH of the turbid solution is adjusted to a value of 6.1 by adding sodium bicarbonate solution. The resulting clear solution was lyophilized to give the pure title product as a pale yellow solid (3.99 g).

¹ H NMR (D ₂ O): δ 2.50-2.70 (m, H), 3.21 (d, 1H, J = 16), 3.30 (t, 2H, J = 8), 3.70 (d, 1H, J = 16 ), 3.78 (s, 2H), 4.04-4.10 (m, 1H), 5.27 (d, 1H, J = 8), 5.80 (d, 1H, J = 8), 7.18 (d, 1H, J = 8) 8.19 (d, 1H, J = 8), 8.23 (s, 1H).

Pharmaceutical Applications and Formulations

The prodrug of the present invention or a physiologically acceptable salt thereof may be administered to the patient as such or in the form of pharmaceutical compositions in which the above compounds are mixed with suitable carriers or excipient (s). Methods of formulation and administration of drugs can be found in Remington's Pharmacological Sciences (Mack Publishing Co., Easton, PA, latest edition).

Administration Methods

Suitable routes of administration may include, but are not limited to, oral, rectal, transmucosal, intramuscular, subcutaneous, intramedicular, intrathecal, intraventricular, intravenous, intravitreal, intraperitoneal, intranasal or intraocular. Preferred routes of administration are oral and intravenous.

In addition, the compound can be administered locally and not by a systemic route, for example, topical administration or by injection directly into the infected part of the patient's body.

Compositions / Compositions

The pharmaceutical compositions in accordance with the present invention can be obtained by methods well known in the art, for example by conventional mixing, dissolving, granulating, preparing dragees, grinding into powder, emulsification, encapsulation, capture or lyophilization processes.

The pharmaceutical compositions can be formulated by conventional methods using one or more physiologically acceptable carriers, including excipients and excipients, which facilitate the incorporation of the active compounds into preparations for pharmaceutical use. The exact composition depends on the chosen route of administration; such formulations are well known to those skilled in the art and include, but are not limited to, the following:

For injection, the compounds may be in aqueous solutions, preferably in physiologically acceptable buffer solutions, such as Hanks solution, Ringer's solution or physiological saline buffer solution. For transmucosal administration, penetrating substances suitable for overcoming the barrier are used in the composition. Such penetrants are well known in the art.

For oral administration, the compounds may be combined with pharmaceutically acceptable carriers well known in the art. Such carriers make it possible to obtain tablets, pills, lozenges, dragees, capsules, solutions, gels, syrups, lotions, suspensions and the like for oral use by the patient based on the compounds of the invention. Pharmaceutical preparations for oral administration can be obtained using a solid excipient, optionally by grinding the resulting mixture and preparing a mixture of granules, after adding other suitable excipients, if necessary, to obtain tablets or dragee inside. Some useful excipients, not limited to, are excipients such as sugars such as lactose, sucrose, mannose or sorbitol, cellulosic components such as corn starch, wheat starch, rice starch and potato starch and other compounds such as gelatin, tragacanth resin , methyl cellulose, hydroxypropyl methyl cellulose, sodium carboxymethyl cellulose and polyvinyl pyrrolidine (PVP). If necessary, loosening agents, such as cross-linked polyvinylpyrrolidone, agar or alginic acid, may be added. Salts such as sodium alginate may also be used.

The inside of the dragee is usually coated with suitable coatings. For this purpose, concentrated sugar solutions may be used, which may optionally contain gum arabic, talc, polyvinyl pyrrolidone, gel cargopol, polyethylene glycol and / or titanium dioxide, lacquer solutions and suitable organic solvents or solvent mixtures. Dyestuffs or pigments may be added to the tablet or dragee coatings to identify or characterize various combinations of doses of the active compound.

Pharmaceutical compositions that can be used orally include suitable capsules made from gelatin, soft, sealed capsules made from gelatin and a plasticizer, such as glycerol or sorbitol, and the like. Capsules suitable for administration may contain the active ingredients in admixture with an excipient, such as lactose, a binder, such as starch, and / or a lubricant, such as talc or magnesium stearate, and, optionally, stabilizers. In soft capsules, the active compounds may be dissolved or suspended in suitable liquids, such as fatty oils, liquid paraffin or liquid polyethylene glycols. Stabilizers may also be added to these formulations.

For inhalation administration, the compounds are usually formulated as an aerosol spray using a pressure-resistant balloon or nebulizer and a suitable nebulizing agent, for example, but not limited to, dichlorodifluoromethane, trichlorofluoromethane, dichlorotetrafluoroethane or carbon dioxide. In the case of using a pressure-resistant aerosol, a single dose can be controlled using a valve to administer a measured amount. Capsules and cartridges, for example, gelatin, for use in an inhaler or blower, may be prepared with a powder mixture of the compound and a suitable base powder, such as lactose or starch.

The compounds may also be formulated for parenteral administration, for example by pill injection or by continuous infusion. Compositions for injection can be in a single dosage form, for example in ampoules or in multidosed containers, with the addition of a preservative. The compositions may take such forms as suspensions, solutions or emulsions in an oily or aqueous binder, and may contain additional compounds, such as suspending, stabilizing and / or dispersing agents.

Pharmaceutical compositions for parenteral administration include aqueous solutions of water-soluble forms, such as, but not limited to, the salt of the active compound. In addition, suspensions of the active compounds can be prepared in a lipophilic binder. Suitable lipophilic binders include fatty oils such as sesame oil, synthetic fatty acid esters such as ethyl oleate and triglycerides, or materials such as liposomes. Aqueous injection suspensions may contain substances, such as sodium carboxymethyl cellulose, sorbitol or dextran, which increase the density of the suspension. Optionally, the suspension may also contain suitable stabilizers and / or agents that increase the solubility of the compounds, resulting in highly concentrated solutions.

In addition, the active ingredient may be in powder form for constitution with a suitable binder, for example sterile pyrogen-free water, before use.

The compounds can also be used in rectal formulations, such as suppositories or delaying enemas, using, for example, conventional suppository bases, such as cocoa butter or other glycerides.

In addition, the compounds can also be used as spare preparations. Such long acting formulations may be administered by injection (eg, subcutaneously or intramuscularly) or by intramuscular injection. The compound can be used for this route of administration with suitable polymeric or hydrophobic materials (for example, in an emulsion with a pharmacologically acceptable oil), with ion exchange resins, or as a sparingly soluble derivative, such as, but not limited to, sparingly soluble salt.

The pharmaceutical compositions may also include suitable solid or gel carriers or excipients. Examples of such carriers or excipients include, without limitation, calcium carbonate, calcium phosphate, various sugars, starches, cellulose derivatives, gelatin and polymers such as polyethylene glycols.

The compounds of the invention may be in the form of physiologically acceptable salts in which the claimed compound may form negatively or positively charged groups. Examples of salts in which the compound forms a positively charged group include, but are not limited to, quaternary ammonium salts such as hydrochloride, sulfate, carbonate, lactate, tartrate, maleate, succinate, where the nitrogen atom of the quaternary ammonium group is the nitrogen of the compound that reacts with suitable acid. Salts in which the compound of the present invention forms a negatively charged group include, but are not limited to, sodium, potassium, calcium, and magnesium salts formed by reacting the carboxyl group of the compound with a suitable base, for example sodium hydroxide (NaOH), potassium hydroxide (KOH), hydroxide calcium (Ca (OH) ₂ ), etc.

A therapeutically effective amount of a β-lactam antibiotic compound of the present invention may be administered to a patient to improve or excrete methicillin, or other β-lactam, such as vancomycin or ampicillin, which are resistant to bacterial infection. In particular, using a therapeutically effective amount of a compound of the present invention, it is possible to cure infectious infections caused by resistant S.aureus types, such as, but not limited to, S.aureus Col (Meth ^R ) (bla-), S.aureus 76 (Meth ^R ) (bla +), S.aureus ATCC 29213, S.Aureus ATCC 25913, S.Aureus ATCC 32432 and S.Aureus CoIBA, or resistant Enterococcus types such as E.faecium ATCC 35667 or E.faecalis ATCC 29212. Of course, the compounds in accordance with the present invention can also be used against bacteria that are susceptible or sensitive to methicillin, vancomycin y, ampicillin or other antibiotics.

Compositions containing a compound or compounds of the invention may be administered for prophylaxis or therapeutic treatment. For therapeutic use, the compositions are administered to an already infected patient, as described above, in an amount sufficient to cure or at least partially eliminate the symptoms of infection. An amount sufficient to achieve this is defined as a “therapeutically effective amount or dose” and depends on the severity and course of the infection prior to treatment, the general condition of the patient and the tolerance of the drugs and the opinion of the attending physician. For prophylactic purposes, compositions containing a compound or compounds of the invention are administered to a patient who has previously been treated with a therapeutically effective amount of the compound to prevent recurrence of infection, or they may be administered to a patient who, for one reason or another, is susceptible to infection but not yet infected, as in the case of surgical administration of antibiotics. Such an amount is defined as a "prophylactically effective amount or dose." The exact amounts also depend on factors such as those described above.

Doses

The exact dose depends on the severity and course of the infection preceding the treatment, the general state of the patient’s health, his or her drug tolerance, etc., all of which are known, the experience and opinions of the attending physician.

Typically, a suitable effective dose of a compound of the invention is in the range of 0.1 to 10,000 milligrams (mg) per patient per day, preferably in the range of 20 to 2000 mg per day. The desired dose is preferably presented in one, two, three, four or more sub-doses, administered at a suitable interval throughout the day. These sub-doses may be administered as unit dosage forms, for example, containing from 5 to 1000 mg, preferably from 10 to 100 mg, of the active ingredient per unit dosage form. Preferably, the compounds of the invention are administered in amounts between about 2.0 mg / kg and 250 mg / kg of the patient’s body weight, from about one to four times a day.

At the first observed improvement in the patient's condition, if necessary, a maintenance dose may be administered during treatment by a doctor. The dose, frequency, or both can be reduced as a function of the condition of the patient to the level at which improvement occurs. When symptoms improve to the desired level, treatment may be discontinued, although some patients may require interruption of treatment for a long period until the outbreak of symptoms resumes.

Biological assessment

The biological activity of compound 1 was previously described as the activity of compound 16 in US 6,025,352, incorporated herein by reference, including any figures, in its entirety. Thus, the biological activity of the prodrugs described herein is expected to be the same, as they are readily hydrolyzed to a related compound in vivo. However, the prodrugs and salts described in the present invention are significantly more water soluble at a neutral or near neutral pH than its salts of a related compound. This is a surprisingly high solubility, which is a major part of the present invention. The solubility of any prodrugs here can be determined using the following method:

A small amount (5-8 mg) of the salt of the compound of the present invention is weighed in a screw cap vial. Water is added to dissolve the salt in high concentration (50-75 mg / ml). Small amounts (1-5 μl increase) of 0.1 N aqueous sodium hydroxide are added to this solution until a slight precipitation of the residue after mixing. The suspension is left at room temperature for 15 minutes, stirring occasionally. An aliquot of the suspension is then transferred to an Eppendorf vial and centrifuged at 14,000 rpm for 1.5 minutes. A portion of the supernatant was dissolved in water, filtered and quantified by HPLC according to a standard calibration curve. The pH of the supernatant is measured using a fine needle sample. For each subsequent determination, using a new sample, add a larger amount of 0.1 N sodium hydroxide for a slight increase in pH, and repeat the remaining procedure. Five to seven determinations are used to obtain a concentration-dependent pH.

In the case of using zwitterion, zwitterion is suspended in water and 0.1 N sodium hydroxide or 0.1 N hydrochloric acid is added until the solid begins to dissolve. Suspensions are left at room temperature for 15 minutes with frequent shaking and then subjected to the same treatment as described above (centrifugation, dissolution and quantification).

An example of a very strong solubility in water at a neutral or near neutral pH of the compounds of the present invention compared with solubility 1 is shown in the drawing. In the drawing, the solubility in water 1 in its zwitterionic form (2) is compared with the solubility of its prodrug L-aspartyl (3):

Determination of the breakdown of prodrugs in serum

Of course, the reason for using the prodrugs of the present invention is their easy conversion to the related active compound 1 under physiological conditions. The ability of a prodrug to turn into 1 can be evaluated using the following method:

Fresh control human serum, fresh control rat serum and monkey rhesus plasma (heparinized and refrigerated) were incubated at 37 ° C. for 15 min in a stirring water bath. serum or plasma pH is measured using a test strip. Then 25 μl of an aliquot of the solution in water of the prodrug of the present invention (2 mg / ml) is added to each group to a final solution volume of 1 ml. Solutions are incubated at 37 ° C for 1 hour, removing 100 μl aliquots 0, 15, 30 and 60 minutes after administration. Aliquots are added to 200 μl of 4% trichloroacetic acid, mixed and centrifuged for 10 min at 14,000 rpm in an Eppendorf microcentrifuge. Then, 25 μl of each supernatant was analyzed using HPLC (Beckman Ultrasphere C 18 column, 5 microns, 4.6 mm × 25 cm and 1 ml / min 95% 0.1 M ammonium acetate, pH = 6.5% acetonitrile, decreasing to 25% acetonitrile in for 20 min with a peak of determination at 254 and 280 nM).

Conclusion

It is well understood by any person skilled in the art that the present invention is well-applied to varying objects, and it is possible to obtain the indicated results and advantages, the same as described in the present description. The molecular complexes and methods described herein, procedures, treatments, molecules, particular compounds, are preferred embodiments, serve as examples, and are not intended to limit the scope of the invention. Substitutions and other uses encountered by a person skilled in the art that fall within the scope of the invention are defined by the scope of the claims.

One skilled in the art will clearly understand that various replacements and modifications may be made in the invention described herein without affecting the scope and breadth of the invention.

All patents and publications mentioned in the description illustrate the level of this technical field to which the invention relates. All patents and publications are hereby incorporated by reference to the same extent as if each specified specific publication were specifically and separately cited as a reference.

The invention, clearly described here, in fact, can be performed in the absence of any element or elements, the limitation or limitations of which are not specifically described here. So, for example, in each example, any of the terms “including”, “consisting of only” and “consisting of” can be replaced by any other two terms. The terms and expressions used are used as terms to describe and not to limit, and the purpose is not to use such terms and expressions to exclude any of the indicated and described equivalents or parts thereof, it is recognized that various modifications are possible within the scope of the claimed invention. It is understood that although the present invention is specifically described by preferred embodiments and optional features, one skilled in the art can make modifications and variations to the concept described herein and that such modifications and variations are included within the scope of the invention as described in the claims.

In addition, where features or aspects of the invention relate to Markush formulas, one skilled in the art will understand that the invention is also described as any individual group or subgroup of members of the Markush formula. For example, if X is selected from the group consisting of bromine, chlorine and iodine, it is fully described that the claimed X may be bromine and X may be bromine and chlorine.

The invention is described here broadly and generally. Each narrower species and subgeneric group included in the generic disclosure also forms part of the invention.

Other embodiments are contained in the following claims.

Claims (15)

1. The compound of the formula

where R ^'1 is hydrogen; R ^{′ 2} represents an acyl group, with the exception of glycyl, which is cleaved by the mammalian enzyme. 2. The compound according to claim 1, in which R ^{′ 2} is selected from the group consisting of —C (O) —R ⁸⁸ , —C (O) —OR ⁸⁹ , —C (O) —CH (NHR ^{′ 3} ) —alk ₄ , and

where r ⁸⁸ is

or

R ^'3 is selected from the group consisting of hydrogen, —C (O) —OR ⁸⁹ and —C (O) —CH (NH ₂ ) —alk ₄ ; alk ₄ is selected from the group consisting of hydrogen and optionally substituted alkyl, where alkyl is optionally substituted with one or more substituents selected from the group consisting of hydrogen, phenyl, —COOH, —C (O) —OR ⁸⁹ , —C ( O) NH ₂ , —OH, —SH, —NH ₂ , and

; R ⁸⁹ is selected from the group consisting of benzhydryl, tert-butyl, allyl, n-nitrobenzyl, benzyl, n- or o-nitrobenzyl, 2,2,2-trichloroethyl, cinnamyl, 2-chloroallyl, tertamyl, trityl, 4-methoxytrityl, 4,4'-dimethoxytrityl, trimethylsilyl, tert-butyldimethylsilyl, β- (trimethylsilyl) ethyl, 4- or 2-methoxybenzyl, 2,4-dimethoxybenzyl, 3,4-dimethoxybenzyl, 2,4,6-trimethoxybenzyl, methoxymethyl and 3.3 dimethylallyl. 3. The compound according to claim 2, in which R ^{′ 3} is selected from the group consisting of hydrogen, methyl, and —C (O) CH (NH ₂ ) CH ₃ . 4. The compound according to claim 2, where alk ₄ is selected from the group consisting of hydrogen, —CH ₃ , —CH (CH ₃ ) ₂ , —CH ₂ OH, —CH ₂ NH ₂ , —CH ₂ CH ₂ NH ₂ , —CH ₂ CH ₂ CH ₂ NH ₂ , -CH ₂ CH ₂ CH ₂ CH ₂ NH ₂ , -CH ₂ COOH, -CH ₂ CH ₂ COOH, -CH ₂ -C (O) NH ₂ , -CH ₂ CH ₂ -C (O) NH ₂ , and

5. The compound according to claim 1, selected from the group consisting of (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ornithylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -prolylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-N- (L) -ananylamino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-aminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L, L) -alanylalanaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -aspartylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N (L) -ananylaminoethylthiomethyl] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) - ( N _α- methyl) alanylaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -histidylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -valaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -aspartylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -lisylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -serylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N (L) -glutaminaminoethylthiomethyl] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, and, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-H - ((5-methyl -1,3-dioxolan-4-en-2-one-4-yl) methoxycarbonyl) aminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid. 6. The compound according to claim 5, selected from the group consisting of (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ornithylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L, L) -alanylalanaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -aspartylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -ananylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -lisylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cephem-4-carboxylic acid, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) -glutamylaminoethylthiomethyl ] pyrid-4-ylthio} -3-cefem-4-carboxylic acid; and, (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-H - ((5-methyl -1,3-dioxolan-4-en-2-one-4-yl) methoxycarbonyl) aminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid. 7. (7R) -7 - [(Z) -2- (2-amino-5-chlorothiazol-4-yl) -2- (hydroxyimino) acetamido] -3- {3- [2-N- (L) aspartylaminoethylthiomethyl] pyrid-4-ylthio} -3-cefem-4-carboxylic acid of the formula:

or a pharmaceutically acceptable salt thereof. 8. A pharmaceutical composition for use in the treatment or prevention of a bacterial infection, containing an effective amount of one or more compounds according to any one of claims 1 to 7. 9. The composition of claim 8, further comprising a pharmaceutically acceptable carrier or excipient. 10. The composition of claim 8 or 9, where the bacterial infection is a bacterial infection that is resistant to β-lactam antibiotic. 11. The composition of claim 10, wherein the bacterial infection resistant to the β-lactam antibiotic is a bacterial infection resistant to methicillin, ampicillin or vancomycin. 12. The composition of claim 10, where the bacterial infection is an infection caused by Staphylococcus or Enterococcus species of bacteria. 13. The composition of claim 12, wherein the Staphylococcus or Enterococcus bacterial species is resistant to one or more other β-lactam antibiotics. 14. The composition of claim 13, wherein the Staphylococcus species are S.aureus Col (Meth ^R ) bla-), S.aureus 76 (Meth ^R (bla +), S.aureus ATCC 29213, S.Aureus ATCC 25913, S.Aureus ATCC 32432 or S. Aureus CoIBA. 15. The composition of claim 13, wherein the Enterococcus species are E.faecium ATCC 35667 or E.faecalis ATCC29212.