NOVEL 3-SUBSTITUTED CEPHEM COMPOUNDS AND PROCESSES FOR PREPARATION THEREOF

Field of the Invention The present invention relates to novel cephalosporins and pharmaceutically acceptable salts thereof, which are useful as antibacterial agents. The invention also relaLes to processes for the preparation of the same, to a pharmaceutical composition containing at least one of the same as active ingredients and to a use of the same for treatment of diseases caused by bacterial infection in human beings and animals.

Background of the Invention

Hereto, it has been known that as antibiotics a great variety of cephalosporins having the basic skeleton of the nucleus of the forraula(A) •

1

(A) have effective antibacterial activities and widely used as antimicrobial agents for infectious diseases caused by various Gram positive and Gram negative strains.

A variety of cephalosporins whose 7-position has 2-arainothiazolyl acetaraino group substituted with α-oxyimino group have been known.

Particularly, the following compounds exhibiting strong antibacterial activities against the enterobacteria have been reported: Cefotaxime in USP 4,152,432 and 4,098,888; Cefmenoxime in USP 4,098,888 and 4,476,122; Ceftriaxone in USP 4,327,210; Ceftazidime in USP 4,258,041 and 4,32&,453.

These compounds, however, show relatively weak antibacterial activities against Staphylococcus when compared with Cephaloridine in French Patent No. 1,384,197 or Cefazoline in USP 3,515,997.

Moreover, Ceftazidime exhibits poor antibacterial activity agains Staphylococcus. although its antibacterial activity against Pseudomonas is relatively stronger than that of other known cephalosporins.

The present inventors have made intensive researches to prepare cephalosporins having improved antibacterial activities against Staphylococcus with strong antibacterial activities against Pseudomonas. As a result of prolonged studies on numerous cephalosporins, we found the 3-substituted cephem compounds show superior antibacterial activities against both of Staphylococcus and Pseudomonas.

Summary of the Invention An object of the invention is to provide novel cephalosporins represented by the formula (I):

wherein,

Ri represents a Cι ~C ₄ alkyl group or - C00R ₄ wherein, R ₂

and R3, independently, represent hydrogen or a Cι~C3 alkyl group and R ₄ represents hydrogen or a Cι~C alkyl group;

Ri* represents hydrogen or an araino-protecting group! represents CH or Ni and the substituent of the formula represents a saturated or unsaturated heterocyclic group containing 1 to 4 nitrogen atoms of which one. nust be substituted with amino group to form quaternary ammonium, and oxygen or sulfur atoms, or a fused heterocyclic group thereof formed together with a substituted or unsubstituted benzene or optional heterocyclic group, and a pharmaceutically acceptable salt thereof.

The other object of the invention is to provide processes for preparing the cephalosporins of the formula (I).

The another object of the invention is to provide pharmaceutical compositions containing at least one compound of cephalosporins as active ingradients.

The another object of the invention is to provide a use of cephalosporins of the formula (I) for treatment of the diseases caused by bacterial infection.

Detailed Description of the Invention

The novel cephalosporins according to the invention may be represented by the following formula (I):

wherein,

Ri represents a Ci~~C ₄ alkyl group or - c - C00R ₄ wherein, R2 and

R3, independently, represent hydrogen or a Cι~C3 alkyl group and R4 represents hydrogen or a Cι~C alkyl group;

Ria represents hydrogen or an amino-protecting group;

Q represents CH or N; and

the formula ^J& represents a saturated or unsaturated

heterocyclic group containing 1 to 4 nitrogen atoms of which one must be substituted with amino group to form quaternary ammonium, and oxygen or sulfur atoms, or a fused heterocyclic group thereof formed together with a substituted or unsubstituted benzene or optional heterocyclic group.

The examples of the 3-position substituent ( γ J may include

1-arainopyrrolidinium methyl, 1-aminopiperidinium methyl, (4-aminomorpholin-4-ium)methyl ,

(1-amino-l,2,3,6-tetrahydropyridinium)methyl , (l-amlno-3-pyrrolin-l-iurn) ethyl , 1-arainoindolinium methyl, 1-arainopyrrolium methyl 1-aminoindolium methyl (l-amino-4-hydroxyρiperidiniua) ethyl, (2-amino-l,2,3,4-tetrahydroisoquinolin-2-iura)methyl, (1-amino-l-raeth 1-4-piperaziniurn)methyl, (4-amino-l,2,4-triazol-2-ium)methyl, (1-amino-l, 2,3-triazol-3-ium)raethyl, (1-amino-l,2,4-triazol-4-ium) ethyl, (l-aminobenzotriazol-3-άum) ethyl, (l-aminobenziaidazol-3-lum)methyl, (l-aminoiraidazol-3-iujι)methyl, (l-aminopyrazol-2-ium)methyl , (l-aminotetrazol-4-ium)methyl ,

(1-amino-l,4.5.6-tetrahydropyriraidin-3-iura)raethyl and (l-aminoindazol-2-ium)methyl.

The C-7 position of the formula (I) may be

represented by - (anti(E)

isomer). However, generally, the syn isomer is preferred.

For the present invention, both of the starting materials and the products are syn isomer.

The pharmaceutically acceptable salts of the compounds (I) are conventional non-toxic salts and may include an inorganic acid salt (e.g., sulfate, hydrochloride, hydrobromide, iodide, phosphate etc.), an

organic carboxylic acid salt (e,g., acetate, maleate, tartrate, fumarate, citrate, succinate, raalate, lactate, oxalate, etc.), a sulfonic acid salt(e.g., ethanesulfonate, benzenesulfonate, p-toluenesulfonate, etc.), a salt with a basic, or acidic amino acid (e.g., salts with arginine, asparagine, glutaraine, lysine, etc.), an inorganic salt, for example, metal salts such as an alkali metal salt (e.g., sodium salt, potassium salt), and an alkaline earth metal salt (e.g. magnesium salt), an ammonium salt, and an organic salt, (e.g., triraethylamine salt, triethylaraine salt, pyridine salt, procaine salt, picoline salt, dicyclohexylamine salt, N-raethylglucamine salt, diethanolamine salt, triethanolamine salt, phenyiethylbenzylamine salt, dibenzylethylenediaraine salt, etc.).

For the present invention, an inorganic acid salt such as sulfate or hydrochloride and an organic acid salt such as maleate or fumarate are particularly preferred since they are formed stable crystalline salts of the cephalosporins(I)

According to the present invention, processes for preparing the cephalosporins(I) and pharmaceutically acceptable salts thereof are provided. The cephalosporins according to the invention may be prepared by the following processes:

The First reaction scheme ^;

(i)

Wherein, Ri and Q are the same as defined above;

Ria represents hydrogen or an araino-protecting group; Bi represents hydrogen or a carboxyl-protecting group;

q represent integer of 0 or l;

the formula is the same as defined above; and the dotted line means ceph-2-em or ceph-3-era compound.

In the first reaction scheme, the compound of the formula (II) as an intermediate may be synthesized by the following procedure:

Wherein,

Bi, q, and the dotted line are the same as defined above; X represents a leaving group, which is halogen or an acetoxy group; and A represents an amine or i ine group which are protected by amino protecting groups. The second reaction scheme:

(I)

Wherein,

Ri, Riβ, , Bi, q, and the dotted line are the same as

defined above; and

W represents a residue which may be substituted with nucleophile.

In preparing the compounds of the formula (I) according to the first or second reaction scheme, any of the following steps can be suitably conducted, if necessary: (i) Removing of carboxyl- or amino- protecting group

(ii) Conversion of Δ ² isomer to Δ ³ isomer, or vice versa

(iii) Conversion(Reduction) of sulfoxide compound (q=l) to sulfide compound (q=0) (iv) Formation of non-toxic salt Preferred compounds of the formula (II) and (VII) in the above first or second reaction scheme are those compounds in which q is 0 and the dotted line represents ceph-3-em.

Δ ² cephalosporin ester derivative (q=l) obtained according to the process of the invention may be converted to Δ ³ cephalosporin ester by treating with base such as triethylamine or pyridine. In case that sulfoxide compound (q=l) is obtained, it may be converted to sulfide compound by treating with reducing agent such as sodium dithionite.

The amino- or carboxyl- protecting group may be suitably selected from the conventional protecting groups which are commonly employed in the technical field of #-lactara and peptide synthesis.

The amino-protecting group may include phthaloyl, formyl, onochloroacetyl, dichloroacetyl, trichloroacetyl, methoxycarbonyl,

ethoxycarbonyl, t-butoxycarbonyl, trichloroethoxycarbonyl, benzy1oxycarbony1, p-ni robenzy1oxycarbony1, di heny1raeth 1oxycarbony1 , methoxyiiethyloxycarbonyl , trityl, trimethylsilyl , phenylacetyl , saiicyl, benzoyl and the like. The carboxyl-protecting group may be include t-butyl, t-amyl, benzy1, p-nitrobenz 1, p-methoxybenzy1 , benzhydr 1, pheny1, p-nitrophen l, raethoxymethyl , ethoxy ethyl , benzyloxyraethyl, acetoxymethyl, raethylthiomethyl, trityl, trichloroethyl, trimethylsilyl, dimethylsilyl, diraethylaminoethyl and the like. The compounds of the formula (II) can be prepared by the method described in Llynn et al., Cephalosporins and penicillin, Academic Press, p 151~171 (1972). For example, they can be prepared by reacting 7 β-acylaraino-3-halomethyl-3-cephem-4-carboxylate derivatives disclosed in Japanese Patent Unexamined Publication Nos. 83-72590 or 83-154588 with the compounds of the formula (IX) or (IXa).

The compounds of the formula (II) also can be prepared by a conventional method using acids, for example, trifluoroacetic acid, formic acid, cone.hydrochloric acid or phosphorus haiide (e.g., phosphorous pentachloride, phosphorus oxychloride, etc.). The acylation in the first reaction scheme is conducted by reacting 1 mole of the co pound(II) with 1 to 3 moles of carboxylic reactive derivative of the compound(III).

The examples of the reactive derivatives may include an acid haiide, an acid anhydride, an. active amide, an active ester and the ¹ like. Preferred examples are an acid haiide such as acid chloride or acid bromide, a mixed acid anhydride such as acetic acid, pivalic acid, isopivalic acid or trichloroacetic acid, an active amide such as

pyrazole, imidazole, dimethylpyrazole or benzotriazole, and an active ester such as p-nitrophenylester, 2.4-dinitrophenylester, trichlorophenylester, l-hydroxy-lH-2-pyridone, N-hydroxysuccinimide or N-hydroxyphthalide. The acylation is preferrably conducted in the presence of condensing agents in case that the compounds(III) are a free acid form. The examples of the condensing agent may include a carbodiimde compound such as N,N-dicyc1ohexy1carbodiimide, N-cyc1ohexy1-N'-morpho1inoeth 1- carbodii ide, N-cyc1ohexyl-N'-(4-diethylarainocyclohexy1)carbodiimide and the like. The acylation also may be conducted in the presence of Vilsmeier reagents formed from the reaction of amide compounds such as N-methylforraaraide or N,N-dimethylforraamide with halides such as thionyl chloride, phosphorus oxychioride or phosgene.

When acid halides or acid anhydrides are used as reactive derivatives, it is necessary to conduct acylation in the presence of acid condensing agents, for example an organic base such as triethyla ine, trimethylamine, ethyldiisopropyla ine, N,N-dimethylaraine, N-methylmorpholine or pyridine, an alkali metal compound such as hydroxide, carbonate or bicarbonate of sodium, potassium or calcium, and oxylan such as ethylene oxide or propylene oxide.

The reaction is carried out in the presence of the solvents which do not influence the reaction adversely. The examples of the solvent may include water, acetone, acetonitrile, dioxane, tetrahydrofuran, dichloromethane, chloroform, dichloroethane, N,N-dimethylforraaiade or mixtures thereof.

The reaction temperature is not specifically limited, but generally ranges between -3CC and 40°C.

. The reaction time generally ranges between 30 min and 10 hours.

The protecting group should be removed in case that the acylated product has a protecting group. The method for removing the protecting group may be suitably selected according to the kind of the protecting group from the methods using an acid, methods using a base or methods using a hydrazine, which are generally used in the field of _/ 9-lactam and peptide synthesis.

With regard to the process of the second reaction scheme, the example of the residue W may include an acetoxy group and a halogen atom such as chlorine, bromine or iodine.

The compounds of (VII) wherein Q is N, Bi is hydrogen and W is an acetoxy group are may be prepared by a method described in The Journal of Antibiotics, Vol. 36(8), 1020(1983).

The reaction of the compounds (VII) wherein W is an acetoxyl group with the compounds (IX) or (IXa) is preferrably carried out in the polar solvents, for example, water, phosphate buffer, acetone, acetonitrile, N,N-dimethylforraaraide, N.N-dimethylacetaraide, tetrahydrofuran, dimethylsulfoxide, dioxane, methanol or ethanol and mixtures thereof with water. The reaction is preferrably carried out around the neutral pH.

The reaction temperature is not specifically limited, but is preferrably 15 ^* C to about 70 ^* C.

The reaction time varies depending on the reaction conditions, but is generally 1 to 10 hours. The reaction may be accelerated by adding alkali metal halides, for example, sodium iodide, lithium iodide or potassium iodide to the reaction mixture.

In case that the compounds (VII) wherein W is a halogen are used to prepare the desired compounds (I), the halogen may include chlorine, bromine and iodine. The haiide compound are easily prepared by methods in Japanese Patent Unexamined Publication Nos. 81-131,590, 83-90,590 and 84-10,593.

The reaction is preferrably carried out in the presence of a solvent such as acetone, dichloromethane, acetonitrile, N,N-dimethylformaraide or N,N-dimethylacetaraide under the non-aqueous condition.

The reaction temperature is preferrably 0 to 5CC and the reaction time is generally 1 to 5 hours.

The protecting group can be removed by known methods to give the compounds(I).

The compounds(IX) or (IXa) which will be introduced into 3-position of the cephe nucleus as substituents are commercially available. For example, l-aralnopyrrolidine, 4-arainomorpholine, 1-aminopiperidine and 4-amino-l,2,4-triazole were purchased from Aldrich. They are also may be synthesized by the known methods. For example, 1-aminopyrrole, 1-aminoindole, 1-amino-l,2,3-triazole, 1-aminobenzotriazole, 1-aminobenzimidazole and 1-aminoindazole were prepared by methods described in Flitsch, W. et al., Chem. Ber., 102. 3268 (1969), Masanori, N. et al., Tetrahedron Lett., 461 (1974), Stolle, R. et al., Chem. Ber., 5£, 1743 (1926), Campbell, CD. et al., J. Chem. Soc. Chem. Commun. , 742 (1969), Sheng, M.N. et al., J. Org. Chem., 28.. 736 (1963) and Sakai, K. et al., J. Org. Chem., 37, 2351 (1972), respectively. Further, for example, l-araino-1,2,3,6-tetrahydropyridine, l-amino-3- pyrroline, 1-aminoindoline, 1-aminohydroxypiperidine, 2-araino-l,2,3,4- tetrahydroisoquinoline, 1-amino-l,2, -triazole, 1-aminoiraidazole,

1-aminopyrazole, 1-aminotetrazole and l-amino-l,4,5,6- tetrahydropyrimidine also may be prepared by the above methods. Alternatively, they also may be prepared by reacting 1,2,3,6-tetrahydropyridine, pyrroline, indoline, hydroxypiperidine, 1,2,3,4-tetrahydroisoquinoline, 1,2,4-triazole, imidazole, pyrazole or tetrazole with hydroxylanine-O-sulfonic acid or chloramine-T in the presence of an inorganic salt in water or a mixed solvent of water and an organic so1vent(l:1^-3:1). If necessary, they may be separated and purified by silica gel column chromatography. The organic solvents are of water-aiscible and may be suitably selected from N,N-dimethylforraaaide, N,N-dimethylformacetamide, methanol, ethanol, acetone, acetonltrile or dioxane.

The inorganic salt may be suitably selected from sodium hydroxide, potassium hydroxide, calcium carbonate or sodium hydride. Particularly, in case that the 3-position substituent is 1- amino-1-methylpiperaziniura, the amino group of 1-methylρiperazine is protected by a conventional araino-protecting-group, for example, acetyl, benzyloxycarbony1 or t-butoxycarbonyl group. Then, thus obtained araino-protected 1-methylpiperazine is reacted with hydroxyl nine-0- sulfonic acid in the presence of an inorganic solvent in water or a mixed solvent of water and an organic solvent to give amino-protected l-araino-l-methylpiperaziniua and the a ino-protecting group is deprotected to give the desired compound.

When the heterocyclic compound having two or nore nitrogen atoms as a hetero atom, for example, 4-amino-l,2, -triazole, l-araino-l,2,3-triazole, 1-araino-l,2,4-triazole, 1-aminopyrazole, 1-arainotetrazole, 1-amino-l,4,5,6-tetrahydropyriraidine,

1-aminobenzotriazole, 1-aminobenziraidazole, 1-aminoindazole or 1-arainoimidazole is introduced into 3-position of the cephem nucleus, the compounds of the forraula(I) may exist in tautomeric forms and such tautoraers are the equivalent and also included within the scope of the invention. Namely, the position of the positive charge varies depending on the state of salt, the kind of solvent, the properties of solution, the temperature, the kind of substituent and the like, as follows:

The salts of the compounds (I) according to the invention can be prepared by dissolving the compounds of the formula (I) in an aqueous solution of inorganic or organic acid, preferrably an aqueous solution containing 1 to 10 equivalents of inorganic or organic acid, stirring

the solution at 0-~-5°C for 5 to 10 hours, then precipitating to give a crystalline salt of compound (I).

The compounds (I) and salts thereof according to the invention are novel cephalosporins and exhibit potent and broad antibacterial activities against a variety of pathogenic bacteria including

Gram-positive and negative strains, particuarly against Staphylococcus and Pseudomonas.

In order to illustrate the pharmaceutical usefulness of the compounds (I) of the invention, the representative examples of compounds (I) were teseted for their antibacterial activities against the standard test strains and the clinically isolated strains.

The antibacterial activity was determined by an Agar dilution method as described below.

That is, the two-fold serial dilutions of the antibacterial compounds(1000xg compound/ml) were prepared and dispersed in Muller Hinton Agar in a petri dish to a concentration of 100~~0.002//g/ral. The culture broths of standard test strains were inoculated on the medium to a concentration 10 ⁷ CFU/ml, and incubated at 37 ^* C for 18 hours. The concentration of compounds(I) at which the strains were not grown was regarded as minimum inhibition concentration (MIC).

The results of antibacterial activity test of the representative examples of the compound (I) against 20 standard test strains shown in terms of MIC were indicated in Table 1.

The MIC of the compound of Example 18 against 254 clinically isolated strains were shown in Table 2.

The representative examples of the compounds (I) according to the invention are as follows:

Compound of Example 1 : (6R,7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2- methoxyiminoacetaraido]-3-(l-aminopyrrolidiniuramethyl)- ceph-3-em-4-carboxy1ate Compound of Example 5 : (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(2-amino-l,2,3,4-tetrahydro- isoquino1in-2-iurn)methy1]-ceph-3-em-4-carboxy1ate Compound of Example 11 (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(1-a ino-l-meth 1-4- piperazinium)raethyl]-ceph-3-era-4-carboxylate Compound of Example 12 : (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(4-amino-l,2,4-triazol-2-iura) methyl]-ceph-3-em-4-carboxylate Compound of Example 18 : (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminobenzotriazol-3-iura) methyll-σeph-3-em-4-carboxylate

Compound of Example 19 (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl )-2- methoxyiminoacetaraido]-3-[(l-aminobenziraidazol-3-iura) methyll-ceph-3-em-4-carboxylate Compound of Example 36 : (6R, 7R)-7-[(Z).-2-(2-aminothiazoi-4-yl)-2- (2-carboxyprop-2-oxyiraino)acetamido]-3-[(4-amino-l,2,4- triazol-2-ium)methyl]-ceph-3-em-4-carboxylate Compound of Example 85 : (6R, 7R)-7-[(Z)-2-(5-amino-l, 2,4- thiadiazol-3-yl)-2-methoxyiminoacetamido]-3-[(l- arainobenzotriazol-3-ium) eth l]-ceph-3-em-4-carboxylate Cefotaxime-Sodium was used as a control.

20 standard test strains used in the present invention are pathogenic strains causing various infections such as urinary tract

Infection, respiratory track infection, dermal soft tissue infection, blood plasma infection, stomach-instestine infection or central nerve system infection, and most of them are j3-lactaaase producing strains. They are shown below: Gram-positive bacteria

1. Streptococcus pyogenes A 308

2. Streptococcus pyogenes A 77

3. Streptococcus faeciura MD 86

4. Staphylococcus aureus SG 511 5. Staphylococcus aureus 285

6. Staphylococcus aureus 503 Gram-negative bacteria

7. Escherichia coli 0 55

8. Escherichia coli DC 0 9. Escherichia coli DC 2

10. Escherichia coli TEM

11. Escherichia coli 1507E

12. Pseudomonas aeruginosa 9027

13. Pseudomonas aeruginosa 1592E 14. Pseudomonas aeruginosa 1771

15. Pseudoaonas aeruginosa 1771M

16. Salmonella typhiraurium

17. Klebsiella oxytoca 1082E

18. Klebsiella aerogenes 1522E 19. Enterobacter cloacae P99

20. Enterobacter cloacae 1321E

Table 1. Antibacterial activity of corapounds(I) against the standard test strains; MIC( tg/ml)

Table 2. Antibacterial activity of the compound of

Example 18 against clinically-isolated strains

As shown in Table 1, the compounds(I) of the invention exhibit broader antibacterial spectrum and more potent antibacterial activity against Gram-negative and positive strains in comparison with Cefotaxime-Sodium. Particularly, the compound of Example 18 exhibits superior activity against _/ 9-lactaaase producing strains such as Pseudomonas aeruginosa and Enterobacter cloacae which have clinical problems in the field of cephem antibiotics as well as show strong activity against Gram-positive strains, for example, Staphylococcus aureus which is resistant to the third-generation cephalosporins.

The compounds of Examples 1, 5, 11, 12, 19 and 36 also exhibited excellent antibacterial activity against Gram-positive strains as well as against Gram-negative strains including Pseudomonas.

Particularly, the compounds of Examples 12, 18, 36 and 85 are

expected to be very useful for treatment of an intractable infection diseases caused by Staphy1ococcus or Pseudomonas.

In order to establish the usefulness in clinical applications of the compounds provided by the invention, the compound of Example 18 was tested for stability against the ;3-lactamase and for antibacterial activity on systemic infection model.

First, the stability against the 3-lactamase was tested by determining the relative(percentage) hydrolysis of the compound of

Example 18, relative to that of Cephaloridine, taking the absolute rate of Cephaloridine as 100.

The yS-lactamases employed in the test were directly isolated from

Enterobacter cloacae P99, E.coli TEM and Citrobacter freundii. and the antibiotics for comparison were Cephaloridine, Cephradine, Cefoperazone and Cefotaxime. Cephaloridine, other know antibiotics and the compound of Example 18 were reacted with each of the enzymes isolated from the strains and the

0D were determined at 260nm, λmax of Cephaloridine. The relative hydrolysis of various compounds were calculated relative to the 0D of

Cephaloridine. The results were shown in- Table 3. Table 3. Relative hydrolysis by jβ -1actamase

The antibacterial activity test on systemic infection model was carried out using the compound of Example 18 as follows: 0.3ml of diluted solution containing lethal dose of strain was administered intraperitoneally to the test animal, and then 5 to 0.078 mg/kg of compound was administered intramuscularly. The PDso was calculated by T probit method and the result was shown in Table 4.

Table 4. Treatment effect on systemic infection model

PDso (mg/kg)

Tested strain (The limit of confidence)* Streptococcus 0.27 pyogenes 77A (0.08- -0.47)

* P < 0.05 Further, in order to illustrate the usefulness of the compounds (I) as a drug, compounds of Exaaples 12, 18 and 36 were tested for acute toxcity by using 4 weeks-old male ICR-mouse as a test aalnal.

The compounds were dissolved in physiological saline and phosphate buffered solution (pH 7.0), or if they were not soluble, suspended in carboxyaethylcellulose or gum Arabic and then mixed with phosphate buffered solution.

The solution or mixture was administered to the tested animals intravenously or subcutaneously and, after 1 week, the number of surviving animals was calculated.

The results of the acute toxcity test were shown in Table 5.

As shown in the Table 5, the LDso of the tested compounds are more than 4000 mg/kg when intravenously administered and more than 6000 rag/kg when subcutaneously administered, so that it was demonstrated that their safty as a drug was very high.

As described above, the compounds according to the present invention exhibit potent and broad antibacterial activity against Gram-positive strains as well as Gram-negative strains including Pseudomonas. while its toxcity is very low.

The compounds of the invention may be administered parenterally for treatment of bacterial infection in human beings in a dose of 50 ~ lOOOmg, preferrably 100-"~500mg per adult, 2~~~λ times a day, preferrably.

The pharmaceutical composition according to the invention contains the compounds (I) as an active ingredient ^' in association with solid or liquid excipients. The excipients may include those one which are commonly employed in the filed of antibiotic pharmaceutics.

The pharmaceutical composition may be formulated into solid formulations, for example, tablet, capsule or powder, or liquid formulations, for example, injection solution, suspension or syrup. The liquid formulations are preferred.

The present invention will be described in detail with reference to the following examples, but is not limited thereto.

Reference Example l: Synthesis of l-araino-l,2,3,6-tetrahydropyridine

To a mixed solvent of water(40ml) and methanol(10ml) was added 11.06g(80ramol) of potassium carbonate, and 3.65ral(40mraol) of 1,2,3,6-tetrahydropyridine and 4.52g(40mmol) of hydroxylamine-O-sulfonic acid were added thereto. The mixture was stirred at room temperature for 4.5 hours and concentrated under the reduced pressure to the half of its original volume. The resulting solid was filtered off and the filtrate was extracted with dichloromethane. The organic layer was treated with anhydrous magnesium sulfate and filtered. The filtrate was concentrated under the reduced pressure and then subjected to chromatography over silica gel using 10% raethanol/dichloromethane as an eluent to give 1.77g of the desired compound(45%). Rf = 0.5 (in 10% raethanol/dichloromethane) NMR (D ₂ 0, 5 ) 2.12—2.48 (m, 2H), 3.20 (t, 2H) 3.42—5.91 (m, 2H)

Reference Example 2- Synthesis of l-araino-3-pyrroline

To 20ml of water was added 0.69g(10mmol) of 3-pyrroline and 1.68g(30mraol) of potassium hydroxide was added thereto under stirring. 1.7g(15ramol) of hydroxylamine-O-sulfonic acid was added while cooling the mixture to 5X or below. The reaction mixture was stirred at 40~ 50 ^* C for 3 hours, cooled and the pH was adjusted to 7~8. Then, it was concentrated under the reduced pressure to remove water. To the residue was added methanol. After stirring for 10 min, the solid was filtered off and the filtrate was dried over anhydrous magnesium sulfate.

Concentration under • the reduced pressure gave 0.4g of the desired compound(17%).

Rf = 0.5 (in 10% raethanol/dichloromethane) 2.72—4.21 (br, 2H, NH ₂ ), 4.54(s, 4H) 5.92 (s, 2H)

Reference Example 3' Synthesis of 1-arainoindoline

In 40ml of water was dissolved 2.76g(20mmol) of potassium carbonate thoroughly, and 10ml of acetonitrile and 2.24ral(20mraol) of indoline were added thereto in this order. After stirring the reaction mixture for 10 min, 2.26g(20mmol) of hydroxylamine-O-sulfonic acid was added thereto. After stirring at room temperature for 1.5 hours, the reaction mixture was concentrated under the reduced pressure. The concentrate was extracted with dlchloromethane and treated with anhydrous magnesium sulfate. After filtration, the filtrate was concentrated under the reduced pressure, and the resulting concentrate was subjected to chromatography over silica gel using dichlororaethane as an eluent. The fractions were concentrated to give 0.99g of the desired compound(37%). Rf = 0.17 (in dichlororaethane) NMR (DMS0-d ₆ , δ )

2.73—3.06 (m, 2H, -CH ₂ -) , 3.25—3.68 (ra, 2H,NH ₂ ) 4.10—6.3 (br, 2H, N-CH ₂ ~) , 6.71—7.01 ( , 2H) 7.02—7.29 (m, 2H)

Reference Example 4= Synthesis of 1-arainoindole

To 50ml of N,N-dimethylforraaraide were added 24.55g(440mmol) of

potassium hydroxide and 3.201g(27.3mmol) of lH-indole, and 6.15g (54.4ramol) of hydroxylamine-O-sulfonic acid was added thereto in small amounts under stirring.

After stirring the reaction mixture at room temperature for 1 hour, 50ml of water was added thereto and it was extracted three times with 100ml of benzene. The extract was washed with water, dried and concentrated under the reduced pressure. Silica gel column chromatography of the residue using 50% dichloromethane/n-hexane as an eluent gave 1.16g of the desired compound(32%). a.p. = 41—41.51C NMR (CDCla, δ )

4.73 (br, 2H, NH ₂ ), 6.37 (d, IH)

7.01—7.65 (m, 5H, phenyl)

Reference Exanple 5: Synthesis of 1-aainopyrrole

To 10.0g(61.7mmol) of N-arainophthalimide was added a solution of 12ml (72.4ramol) of 2,5-diethoxytetrahydrofuran in 100ml of dioxane and 10ml of 5N hydrohloric acid was added thereto. The reaction mixture was stirred for 1 hour and cooled to 5'C - The resulting precipitate was filtered, recrystallized from dioxane:waterd:3), washed with ethanol and dried to give 10.3g of 1-phthalimidopyrrole. This product was dissolved in 60ml of methanol and 3ml of 82% hydrazine hydrate was slowly added thereto. After refluxing the reaction mixture for 30 min, cooled to 0—5°C and 1.5ral of glacial acetic acid was added thereto. The resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure to remove methanol. The concentrate was washed with diethyl ether and vacuum-distilled to give

2.4g of the desired compound(47.3%). bp = 71—73^ (12ramHg) NMR (CDCla, δ )

4.5- (s, 2H, NH ₂ ), 5.89 (t, 2H) 6.40 (t, 2H)

Reference Exaraple 6: Synthesis of l-amino-4-hydroxypiperidine

In 10ml of water was dissolved 0.6g(6mmol) of hydroxylamine-O- sulfonic acid and 1.82g(1.8mmol) of 4-hydroxypiperidine was added thereto. After refluxing for 1 hour, the reaction mixture was cooled to 5 2. and 0.84g(6.1mmol) of potassium carbonate was added thereto. The reaction mixture was stirred for lOrain and the precipitate was filtered off. The filtrate was concentrated under the reduced pressure and 15ml of dry ethanol was added to give precipitate, which was filtered off. While cooling the filtrate to 5—10 ^* 0, 0.85ml of 57% HI was slowly added and the mixture was stirred at the same temperature for 30 min. To the reaction mixture was added 50ml of petroleum ether. The filtrate was washed with acetone to give 1.41g of the desired compound(34%). m.p. = 117—1191C

1.12—1.98 (m, 4H), 2.41—2.86 (m, 2H) 2.86—3.24 (m, 2H) , 3.40—3.79 (m, IH) 5.23 (br, 2H, NH ₂ )

Reference Exaraple 7: Synthesis of 2-amino-l,2,3,4-tetrahydroisoquinoline In a mixed solvent of 50ml of water and 20ml of methanol was

dissolved 11.06g (δOmraol) of potassium carbonate and 5.33g(40mmol) of lH-l,2,3,4-tetrahydroisoquinoline and 4.52g(40mmol) of hydroxylaraine-O- sulfonic acid were added thereto while maintaining the temperature at 30 ^* C. The mixture was stirred at room temperature for 4 hours. After completion of reaction, the insoluble was filtered off and the filtrate was extracted twice with dlchloromethane. The dlchloromethane layer was dried over anhydrous aagnesiua sulfate and concentrated to driness. The resulting residue was subjected to silica gel column chromatography using 5% aethanol/dichloromethane as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and crystallized from tetrahydrofuran to give 1.78g of the desired compound in a forn of white crystal(30%) . m.p. = 179—181 ^* C NMR (DMS0-dύ + 10% DC1, δ) 2.95—3.25 ( , 2H, -CH ₂ -), 3.34—3.64 (n, 2H, N-CH2-) 4.36 (s, 2H), 7.04—7.32 ( , 4H, phenyl)

Reference Exaraple 8: Synthesis of 1-aninoimidazole • p-toluenesulfonic acid In 50ml of water was dissolved 2.76g(20amol) of potassium carbonate and 1.36g(20mraol) of imidazole was added thereto. 2.26g(20mraol) of hydroxylaraine-O-sulfonic acid was added thereto and the mixture was stirred at room temperature for 6 hours. The resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure. To the concentrate were slowly added 10ml of chloroform and a solution of 3.8g(20mmol) of p-toluenesulfonic acid • nonohydrate in 10ml of chloroform. Stirring for 30 rain, filtration and drying of the

mixture gave 1.74g of the desired compound(34%). NMR (DMS0-d6, δ )

2.30 (s, 3H, CH ₃ ), 7.26 (m, 4H, phenyl)

7.69 (d, 2H), 9.10 (t, IH)

Reference Exaraple 9: Synthesis of 1-aminopyrazole • p-toluenesulfonic acid In 500ml of water were dissolved 2.76g(20mmol) of potassium carbonate and 1.31g(20ramol) of pyrazole and 2.26g(20mmol) of hydroxylaraine-O-sulfonic acid was added thereto. The reaction mixture was stirred at room temperature for 6 hours and concentrated under the reduced pressure. To the concentrate was added methanol to give precipitate, which was filtered off. After concentrating under the reduced pressure, the concentrate was treated in the same way as- Reference Exaraple 8 to give 1.54g of the desired compound(30%) .

2.32 (s, 3H, CH ₃ ), 6.7 (t, IH) 7.3 (m, 4H, phenyl), 7.65 (d, 2H)

Reference Exaraple 10: Synthesis of 1-aminobenzotriazole

In 200ml of water were dissolved 23.83g(0.2raol) of lH-benzotriazole and 56.llg(lmol) of potassium hydroxide and the solution was maintained at 60 ^* C. To the reaction solution was slowly added 45.2g(0.4mol) of hydroxylanine-0-sulfonic acid over 1 hour while maintaining the temperature at 70—75 ⁰ C. After completion of addition, the reaction mixture was stirred at 70°C for further 1 hour. After completion of reaction, the reaction solution was cooled to room temperature to

precipitate potassium sulfate as crystal, which was filtered off. The filtrate was extracted with dichlororaethane (100ml x 5) and the extract was dried over anhydrous magnesium sulfate and concentrated under the reduced pressure to give yellow crystal. Thus obtained crystal was dissolved in 5ml of tetrahydrofuran and the pH was adjusted to 1 with cone.hydrochloric acid to give white crystal.

The filtered crystal was dissolved in 50ml of water and the pH was adjusted to 11—12 with 30% sodiun hydroxide followed by extraction with dichloromethane(30mlx4). The extract was dried over anhydrous magnesium sulfate and concentrated under the reduced pressure to give

12.07g of the desired compound(45%). m.p. = 83—841C

NMR (CDCla. δ ) 5.80 (br, 2H, NH2), 7.25—7.55 (m, 2H) 7.70—8.11 (m, 2H)

Reference Example 11: Synthesis of 1-amino-l,2,4-triazole

In 20ml of ethanol was dissolved 14.03g(250mmol) of potassium hydroxide and 3.45g(50mmol) of 1,2,4-triazole was added thereto. The solution of 11.31g(100ramol) of hydroxylamine-O-sulfonic acid in a mixed solvent of water(lθml) and ethanol (10ml) was slowly added to the mixture over 30 min. After stirring at room temperature for 2.5 hours, the resulting precipitate was filtered off and the filtra i was concentrated. To the concentrate was added tetrahydrofuran and the mixture was stirred for 30 rain.

The tetrahydrofuran layer was separated, concentrated under the

32

reduced pressure and diethyl ether was added thereto. The mixture was stirred for 30 min and the diethyl ether layer was separated and concentrated under the reduced pressure. Crystallization of the concentrate from diisopropyl ether/n-hexane gave the 1.63g of the desired compound in a form of crystal(46%). m.p. = 48—50"C

8.58 (s, IH), 9.36(s, IH)

Reference Example 12: Synthesis of 1-aminoindazole

To a solution of 2.2g(55ramol) of sodium hydroxide in 30ml of water was added 1.33g(10mmol) of lH-indazole and ethanol was slowly added at 50 ^* C until the reaction mixture was dissolved thoroughly. The resulting mixture was heated to 55 ^* C and 2.83g(25mraol) of- hydroxylamine-O-sulfonic acid was slowly added over 30 rain witli vigorous stirring followed by further stirring for 30 min. After completion of reaction, the resulting precipitate was filtered off, and the filtrate was extracted with dichloromethane(30mlx2), dried over anhydrous magnesium sulfate and concetrated under the reduced pressure. The concentrate was chroraatographed over silica gel using 10% ethyl acetate/dichloromethane as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and crystallized from benzene/petroleum ether to give 0.63g of the desired compound(47%) . m.p. = 104—lt>6°C

5.41 (s, 2H, NH ₂ ), 6.85—7.95 , 4H)

8.05 (s, lH)

Reference Exaraple 13: Synthesis of 1-aminotetrazole

In 125ral of water was dissolved 13.δgdOOmmol) of pσtassiura carbonate and 7g(100mraol) of lH-tetrazole was added thereto. The reaction solution was heated to 75 ^* C and 11.3g(100mmol) of hydroxylaraine-O-sulfonic acid was slowly added at 70— 5°C over 30 min while adjusting its pH to 7—8. After completion of addition, the solution was refluxed for 30—35 rain, the pH was adjusted to 8 and the solution was extracted with ethyl acetate. The extract was concentrated under the reduced pressure to give 1.7g of the desired compound(20%). b.p. = 153—158T 2ramHg) 7.10 (br, 2H, NH ₂ ), 9.12 (s, IH)

Reference Exaraple 14: Synthesis of 1-amino-l,2,3-triazole

In 20ml of distilled water were dissolved 2.76g(20mraol) of potassium carbonate and 1.38g(-20mraol) of 1H-1,2,3-triazole and 2.26g(20mmol) of hydroxylaraine-O-sulfonic acid was added thereto. The mixture was stirred at room temperature for 9 hours, concentrated under the reduced pressure and ethanol was added to give a precipitate, which was filtered off. The filtrate was concentrated under the reduced pressure and crystallized from tetrahydrofuran/diethyl ether/chloroforn to give- 0.96g of the desired product(57%). m.p. = 51 ^* C NMR (DMS0-d6 + D ₂ 0, δ )

7. 8 (d, 2H)

Reference Exaraple 15: Synthesis of 1-aminobenziraidazole

In a mixed slovent of water(δθml) and ethanol(50ml) was dissolved 19.64g(35mmol) of potassium hydroxide and 5.91g(5ramol) of lH-benzimidazole was added thereto. Water(lθral) containing 14.41g (125mraol) of hydroxylamine-O-sulfonic acid was slowly added while maintaining the temperature at 30°C and the mixture was stirred at the same temperature for 18 hours. The insoluble was filtered off, and the filtrate was extracted with ethyl acetate. The ethyl acetate layer was dried over anhydrous magnesium sulfate and concentrated to driness. The concentrate was subjected to column chromatography over silica gel using 50% ethyl acetate/dichloromethane as an eluent. The fractions containing the desired product were concentrated under the reduced pressure to give 3.06g of the desired compound in a form of white crystal (46%). m.p. = 141O NMR (DMS0-d«s, δ)

6.19 (br, 2H, NH ₂ ), 7.08—7.44 (m, 2H) 7.45—7.76 (m, 2H) , 8.13 (s, IH)

Reference Example 16: Synthesis of 1-amino-l,4,5,6-tetrahydropyrimidine

To 2.76g(20ramol) of potassium carbonate were added 30ml of water and

10ml of ethanol, and 1.64ml (20mmol) of lH-l,4,5,6-tetrahydropyrimidine was added thereto. ater(lθml) containing 2.26g(20mmol) of hydroxylamine-O-sulfonic acid was slowly added to the mixture at 301C or below and the resulting mixture was stirred at room temperature for 24

hours. After completion of the reaction, the insoluble was filtered off, and the filtrate was extracted twice with ethyl acetate. The ethyl acetate layer containing the desired product was dried over anhydrous magnesium sulfate, concentrated under the reduced pressure and purified by silica gel column chromatography (eluent: 10% aqueous methanol) to give 0.56g of the desired coapound(28%) . NMR (DMSO-dd, δ)

1.4—1.8 (q, 2H), 3.3—3.8 ( , 4H)

3.6 (s, IH), 6.5 (br, 2H)

Reference Exaraple 17: Synthesis of 1-amino-l-methyl piperaziniua chloride hydrochloride In 30ml of water was dissolved 2.84g(20mβol) of 4-acetyl-l-methyl piperazine and 3.4g(30ramol) of hydroxylamine-O-sulfonic acid was added thereto. To the mixture, 2.4g(60mmol) of sodium hydroxide was added in small amounts and the resulting mixture was stirred at 40 ^* C for 2 hours, cooled to 5 ^* C and the pH was adjusted to 7—8 with cone.hydrochloric acid while keeping the above temperature. The reaction solution was concentrated under the reduced pressure to remove water and methanol was added to the residue. The mixture was stirred for 1 hour and filtered. The filtrate was dried over anhydrous magnesium sulfate and concentrated under the reduced pressure to give 2.8g of l-methyl-l-amino-4-acetylpiperazinium in a form of white crystal (88.5%).

Thus obtained product 2.8g(18amol) was dissolved in 50ml of water and 2.88g (72ramol) of sodium hydroxide was added thereto.

The reaction mixture was stirred at 50X2 for 2 hours and the pH was adjusted to 1 with cone.hydrochloric acid. The mixture was

concentrated under the reduced pressure and methanol was added thereto. The resulting mixture was stirred for 1 hour and the precipitate was filtered off. The methanol solution containing the desired product was dried over anhydrous magnesium sulfate and concentrated under the reduced pressure to give 2.33g of the desired compound in a form of white crystal(62%). m.p. = 230 ^* C (dec.)

3.98—3.65 (m, 4H, -CH2-NH-CH2), 3.53 (s, 3H, CH3) 3.75—4.10 (m, 4H), 6.57 (s, 2H, NH ₂ )

Exaraple 1

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-(l-arainopyrrolidiniuramethyl)- ceph-3-em-4-carboxylate

In 40ml of water was dissolved 8.99g(60mmol) of sodium iodide and the solution was heated to 65—701C. Sodium (6R,7R)-7-[(Z)-2-(2- arainothiazol-4-yl)-2-raethoxyirainoacetamido]-3-acetoxymeth yl-ceph-3-em-4- carboxylate(9.54g,20mraol) and 2.94g(24mmol) of 1-aminopyrrolidine hydrochloride were added thereto and the mixture was stirred for 1.5 hours while adjusting its pH to 6—6.5. The reaction solution was cooled to room temperature, its pH was adjusted to 1—1.5 with 3N hydrochloric acid and then was stirred for 30 min. The resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure and a small amount of water was added thereto. Column chromatography over aluminium oxide(80% aqueous acetonitrile), silica

gel(80% aqueous acetonitrile) and then DIAI0N ® HP-20(Mitsubishi) (80%

aqueous acetonitrile) were conducted. Concentration followed by lyophilization gave 2.12g of the desired product in a form of white power(22%).

Rf = 0.31 (in 80% aqueous acetonitrile) m.p. = 182'C—(decorap.) MS (FAB, M+l) = 482 NMR (20% DC1, δ )

1.75—2.30 (m, 4H) , 2.90—3.70 ( , 6H)

3.80 (s, 3H, 0CH ₃ ), 3.80—4.20 (m, IH, C-3) 4.4—4.9 (m, IH, C-3), 5.17 (dd, IH, C-6)

5.7(dd, IH, C-7), 7.15 (s, IH, thiazole-H)

Example 2

Synthesis of (6R, 7R)-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(4-aminomorphoIin-4-ium) methy1]-ceph-3-em-4-carboxy1ate

By following the procedure in Example 1- except that 2.31ml (24ramol) of 4-aminomorpholine was employed in place of 1-aminopyrrolidine, there was obtained 2.5g of the desired compound( 5%) . Rf •= 0.35 (in 80% aqueous acetonitrile) m.p. = 126 ^* C-(decorap.) NMR (20% DC], δ )

3.35—4.55 (m, 10H, morpholine, C-2), 4.09 (s, 3H, OCH3) 4.95—5.34 (q, 2H, C-3), 5.32 (d, IH, C-6) 5.85(d, IH, C-7), 7.20 (s, IH, thiazole-H)

Exarapl e 3

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-(l-arainopiperidiniumraethyl)- ceph-3-em-4-carboxylate By following the procedure in Example 1 except that 2.59ml(24mmol) of 1-aminopiperidine was employed in place of 1-aminopyrrolidine, there was obtained 2.3g of the desired product(23%). Rf = 0.35 (in 80% aqueous acetonitrile) m.p. = 120'C—(decorap.) NMR (20% DC1, δ)

1.5—2.2 (m, 6H), 3.75 (s, 3H, OCHs), 3.2—4.35 ( , 6H)

4.89—5.2 (m, 2H, C-3), 5.37 (d, IH, C-6)

5.95(d, IH, C-7), 7.18 (s, IH, thiazole-H)

Example 4

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-amino-4-hydroxypiperidiniura) methyl]-ceph-3-em-4-carboxy1ate

By following the procedure in Exaraple 1 except that 2.79g(24mraol) of l-araino-4-hydroxypiperidine was employed in place of 1-aminopyrrolidine, there was obtained 2.1g of the desired product(20%). Rf = 0.15 (in 80% aqueous acetonitrile) NMR (20% DC1, δ )

1.12—1.98 (m, 4H), 3.2—4.4 (m, 7H) 3.85 (s, 3H, OCH3), 4.86—5.2 (br, 2H, C-3) 5.34(d, IH, C-6), 5.96 (d, IH, C-7) 7.2 (s, IH, thiazole-H)

Exaraple 5

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(2-amino-l,2,3,4-tetrahydroisoquin olin-

2-ium)methyl]-ceph-3-em-4-carboxylate In a mixed solvent of acetonitrile(5ml) and water(30ral) was dissolved 4.02g(30nmol) of lithium iodide, 0.99g(6.68ramol) of

2-araino-l,2,3,4-tetrahydroisoquinoline was added thereto and the temperature was raised to 67— 0°C. To the reaction solution was added

1.43g(3ramol) of sodium(6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxy- irainoacetamido]-3-acetoxyraethyl-ceph-3-em-4-carboxylate, and the reaction mixture was stirred at 70—72 ^* C for 2 hours while adjusting the pH to 6.5—7.0.

After completion of reaction, the reaction solution was cooled to room temperature and stirred for 30 min while adjusting the pH to 1—1.5 with IN hydrochloric acid. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure. The concentrate was subjected to column chromatography over aluminium oxide and silica gel(eluent: 80% aqueous acetonitrile).

The fractions obtained were concentrated under the reduced pressure, and the concentrate was dissolved in a small amount of water followed by

purification by DIAI0N ® HP-20(Mitsubishi) column chromatograph (eluent:

8% aqueous ethanol).

The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.23g of the desired compound in a pale yellow amorphous form(14%).

Rf = 0,4 (in 80% aqueous acetonitrile)

MS(FAB, M+l) = 544

NMR (DMS0-d<s + 20% DC1, δ )

3.1—3.5 (m, 2H), 3.5 —4.3 (m, 6H) 4.04 (s, 3H, 0CH ₃ ), 4.5—5.1 (m, 2H, C-3) 5.2(dd, IH, C-6), 5.8 (dd, IH, C-7) 7.09 (s, IH, thiazole-H), 7.1—7.8 (m, 4H, phenyl)

Example 6

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazoi-4-yl)-2- methoxyiminoacetaraido]-3-(l-arainoindoliniummethyl)-ceph- 3-em-4-carboxylate

By following the procedure in Example 5 except that 0.9g(6.68ramol) of 1-aminoindoline was employed in place of 2-araino-l,2,3,4- tetrahydroisoquinoline, there was obtained 0.2g of the desired product (13%). Rf = 0.37 (in 80% aqueous acetonitrile) NMR (DMSO-dώ + 20% DC1, δ)

2.8—3.1 (m, 2H), 3.4 —4.4 (m, 4H) 4.01 (s, 3H, OCH3), 4.6—5.1 (m, 2H, C-3) 5.1(d, IH, C-6), 5.7 (d, IH, C-7) 7.01 (s, IH, thiazole-H), 7.03—7.7 (ra, 4H, phenyl)

Exaraple 7

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-(l-aminoindoliumraethyl)- ceph-3-em-4-carboxylate

By following the procedure in Example 5 except that 0.9g(6.68mmol) of 1-aminoindole was employed in place of 2-amino-l,2,3,4-

tetrahydroisoquinoline, there was obtained 0.26g of the desired product(17%).

Rf = 0.36 (in 80% aqueous acetonitrile)

NMR (DMSO-dβ + 20% DC1, δ ) 3.5—3.9 (m, 2H, C-2), 3.86 (s, 0CH ₃ ) 4.6—5.1 (m, 2H, C-3), 5.2 (d, IH, C-6) 5.6(d, IH, C-7), 6.45 (d, IH), 7.01 (s, IH, thiazole-H) 7.05—7.8 (m, 5H, ^' phenyl)

Exaraple 8

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(1-amino-l,2,3,6-tetrahydropyridi nium) methy11-ceph-3-em-4-carboxy1ate

In a mixed solvent of acetonitrile(4ml) and water(9ral) was dissolved 3.21g(24mraol) of lithium iodide and the solution was heated to 70 ^* .

0.33g(3.36mraol) of 1-amino-l,2,3,6-tetrahydropyridine was added thereto and 0.764g(1.6mmol) of sodium (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- methoxyiminoacetaraido]-3-acetoxymethyl-ceph-3-era-4-carboxy late was added while adjusting the pH to 6.5—7. The mixture was stirred at 70—72*C for 1 hour while adjusting the pH to 6.5—7.0.

The reaction solution was cooled to room teraperature, and its pH was adjusted to 1—-1.5 with 3N hydrochloric acid. The solution was stirred for 30 min, the resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure. The concentrate was dissolved in a small amount of water and subjected to column chromatography over aluminium oxide(80% aqueous acetonitrile), silica

gel(85% aqueous acetonitrile) and DIAI0N ®HP-20 (Mitsubishi) (80% aqueous

acetonitrile). The fractions containing the desired product were concentrated and freeze-dried to give 158rag of the desired compound(20%) .

Rf = 0.29 (in 80% aqueous acetonitrile)

2.0—2.4 (m, 2H), 3.0 —3.9 (m, 6H)

3.97 (s, 3H, OCHs), 4.0—4.3 (m, IH, C-3)

4.7—5.1 (m, IH, C-3), 5.15 (d, IH, C-6)

5.5—5.9 (m, 3H, C-7), 6.73 (s, IH, thiazole-H) 9.5 (d, IH, -NH)

Exaraple 9

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(l-amino-3-pyrrolin-l-ium)methyl] -ceph-3-era-4-carboxylate

By following the procedure in Example 8 except that 0.28g(3.36 ramol) of l-amino-3-pyrroline was employed in place of 1-amino-l,2,3-6- tetrahydropyridine, there was obtained 0.17g of the desired compound (22%). Rf = 0.3 (in 80% aqueous acetonitrile) NMR (DMS0-dώ + 20% DC1, δ )

3.6—4.0 (m, 2H, C-2) , 3.86 (s, 3H, OCH3) 4.6 (s, 4H), 4.56—5.1 (m, 2H, C-3) 5.2(d, IH, C-6), 5.6 (d, IH, C-7) 5.9 (s, 2H), 7.01 (s, IH, thiazole-H)

Exaraple 10

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-(l-aminopyrroliummethyl)- ceph-3-em-4-carboxylate By following the procedure in Example 8 except that 0.28g(3.36ramol) of 1-arainopyrrole was employed in place of 1-amino-l,2,3,6- tetrahydropyridine, there was obtained 0.16g of the desired compound(21%).

Rf = 0.29 (in 80% aqueous acetonitrile) NMR (DMSO-dώ + 20% DC1, δ)

3.6—4.1 ( , 2H, C-2), 3.83 (s, 3H, 0CH ₃ ) 4.6—5.1 (m, 2H, C-3), 5.15 (d, IH, C-6) 5.62(d, IH, C-7), 6.1 (t, 2H) 6.5 (t, 2H), 7.01 (s, IH, thiazole-H)

Exaraple 11

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-amino-l-nethy1-4-piperaziniurn )methyl]- ceph-3-em-4-carboxylate In 8ml of water was dissolved 2.5g(17mmol) of sodium iodide, 8al of acetonitrile was added thereto and the mixture was heated to 65—70°C. 0.376g(2mraσl) of 1-amino-l-raethylpiperazinium chloride hydrochloride and 0.48g(lramol) of sodium (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-acetoxymethyl-ceph-3-era-4-carboxy late were added in this order and the pH was adjusted to 6.8 with sodium bicarbonate solution. The reaction solution was stirred at 65—70°C for 2 hours, cooled to room temperature and washed three times with 50ml

44

of dlchloromethane. The aqueous layer was adjusted to pH 2 with 6N hydrochloric acid, concentrated under the reduced pressure and the resulting precipitate was filtered off. The filtrate was concentrated under the reduced pressure and the concentrate was subjected to silica gel column chromatography (80% aqueous acetonitrile). The fractions containing the desired product were purified by HP-20 column chromatography using the same eluent to give 0.37g of the desired compound(73%).

Rf = 0.11 (in 80% aqueous acetonitrile) m.p. = 194 ^* C—(decorap.) MS(FAB, M+l) = 511 NMR (DMS0-d ₆ , δ)

3.0—3.7 (m, 15H), 3.83 (s, 3H, 0CH ₃ )

4.96 (d, IH, C-6), 5.51 (dd, IH, C-7) 6.03(s, 2H, NH2), 6.73 (s, IH, thiazole-H)

7.18 (br, 2H), 9.45 (d, IH, -NH)

Example 12

Synthesis of (6R,7R)-7-t(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(4-amino-l,2,4-triazol-2-iura)raet hyl]- ceρh-3-em-4-carboxylate In 25ml of distilled water was dissolved 4.3g(30mmol) of sodium iodide and the solution was heated to 70—-75'C. 1.0g(12mmol) of 4-amino-l,2,4-triazole and 4.77g(10mraol) of sodium (6R,7R)-7-[(Z)- 2-(2-aminothiazol-4-yl)-2-methoxyirainoacetamido]-3-acetoxyr aethyl-ceph-3- em-4-carboxylate were added thereto and the pH was adjusted to 6-—6.5. The reaction mixture was stirred at 70—75°C for 1 hour, the reaction

solution was cooled to room temperature and the pH was adjusted to 1—-1.5 with 3N hydrochloric .acid. The resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure. The concentrate was dissolved in a small amount of water and subjected to column chromatography over aluminum oxide(80% aqueous acetonitrile) and HP-20 (85% aqueous acetonitrile). The fractions containing the desired product were concentrated and freeze-dried to give 1.68g of the desired product(35%). Rf = 0.23 (in 80% aqueous acetonitrile) m.p. = 170 ^β C-(decorap.) MS(FAB, M+l) = 480

3.3—3.6 (m, 2H, C-2), 3.83 (s, 3H, 0CH ₃ ) 5.04 (d, IH, C-6), 5.0—5.35 (q, 2H, C-3) 5.6(d, IH, C-7), 7.01 (s, IH, thiazole-H) 8.76, 9.81 (s,s, 2H, triazσle)

Exaraple 13

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(1-amino-l,2,4-triazol-4-iun) methyl]-ceph-3-em-4-carboxylate

In a nixed solvent of acetonitrile (10ml) and water(lθ l) was dissolved 9.69g(64.65 raol ) of sodium iodide and the solution was heated to 70—75 ^* C. 1.63g(19.4ramol) of 1-araino-l,2,4-triazole and 6.17g(12.93 mraol) of "sodium (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxy- i11inoacetamido]-3-acetoxyraeth l-ceph-3-em-4-carboxylate were added thereto, and the raixture was stirred at 70—75 ^* C for 1.5 hours while

adjusting the pH to 6—6.5 and cooled to room temperature. After adjusting the pH to 1—1.5 with 3N hydrochloric acid, the reaction mixture was stirred for 30 rain. The resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure. The concentrate was dissolved in a small amount of water and subjected to column chromatography over aluminium oxide (80% aqueous acetonitrile), silica gel (85% aqueous acetonitrile) and then HP-20 (Mitsubishi) (80% aqueous acetonitrile). The fractions containing the desired product were concentrated and freeze-dried to give 2.05g of the desired compound (33%).

Rf = 0.31 (in 80% aqueous acetonitrile) m.p. = 166'C-(decorap.) MS(FAB, M+l) = 480 NMR (DMSO-dή + 20% DC1, δ) 3.28—3.7 (ra, 2H, C-2), 3.83 (s, 3H, 0CH ₃ ) 5.1 (q, 2H, C-3), 5.18 (d, IH, C-6) 5.61(d, IH, C-7), 6.91 (s, IH, thiazole-H) 8.74, 9.68(s,s, 2H, triazole)

Exaraple 14

Syhthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(l-aminoiraidazol-3-iura)raethy1]- ceph-3-em-4-carbox late

By following the procedure in Exaraple 13 except that 1.61g(19.4ramol) of 1-aminoimidazole was employed in place of 1-amino-l,2,4-triazole, there was obtained 1.95g of the desired compound(32%) . Rf = 0.29 (in 80% aqueous acetonitrile)

NMR (DMSO-dώ + 20% DCl , δ )

3.2—3.74 (ra, 2H, C-2) , 3.86 (s, 3H, 0CH ₃ ) 5.08 (q, 2H, C-3), 5.2 (d, IH, C-6) 5.6(d, IH, C-7), 6.95 (s, IH, thiazole-H) 7.6, 8.3 (s,s, 2H), 9.3 (s, IH)

Example 15

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminopyrazol-2-ium)methyl]- ceph-3-em-4-carboxylate

By following the procedure in Example 13 except that 1.61g(19.4mmol) of 1-aminopyrazole was employed in place of 1-amino-l,2,4-triazole, there was obtained 1.81g of the desired compound(29%). Rf = 0.3 (in 80% aqueous acetonitrile) NMR (DMSO-de + 20% DCl, δ )

3.27—3.74 (m, 2H, C-2), 3.9 (s, 3H, OCH3)

5.1 (q, 2H, C-3), 5.15 (d, IH, C-6) 5.68(d, IH, C-7), 6.7 (t, IH)

7.02 (s, IH, thiazole), 7.8, 8.4 (s,s 2H)

Exaraple 16

Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminotetrazol-4-iura)methyI]- ceph-3-em-4-carboxylate By following the procedure in Exmaple 13 except that 1.67g(19.4ramol) of 1-aminotetrazole was employed in place of 1-amino-l,2,4-triazole, there was obtained 1.73g of the desired compound(28%).

Rf = 0.25 (in 80% aqueous acetonitrile) NMR (DMSO-da + 20% DCl, δ)

3.20—3.74 (m, 2H, C-2), 3.83 (s, 3H, 0CH ₃ )

5.12 (q, 2H, C-3), 5.21 (d, IH, C-6) 5.63(d, IH, C-7), 7.01 (s, IH, thiazole-H)

9.7 (s, IH, tetrazole)

Exaraple 17

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(1-amino-l,2,3-triazol-3-iura) methyl]-ceph-3-em-4-carboxylate

By following the procedure in Exaraple 13 except that 1.63g(19.4ramol) of 1-araino-l,2,3-triazole was employed in place of 1-amino-l,2,4- triazole, there was obtained 2.02g of the desired compound(33%). Rf = 0.24 (in 80% aqueous acetonitrile) m.p. = 126'C —(decomp.) MS(FAB, M+l) = 480 NMR (DMSO-dβ + 20% DCl, δ )

3.21—3.68 (m, 2H, C-2), 3.9(s, 3H, 0CH ₃ ) 5.12 (q, 2H, C-3), 5.16 (d, IH, C-6)

5.65(d, IH, C-7), 7.01 (s, IH, thiazole-H) 7.8, 8.7 (s,s, 2H, triazole)

Exaraple 18 Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(l-aminobenzotriazol-3-iura) ethy1]-ceph-3-em-4-carboxy1ate

In a mixed solvent of acetonitrile(lθml) and water(15ml) was dissolved 6.69g(50mraol) of lithium iodide and the solution was heated to 65—67 ^* C. 2.42g(18mmol) of 1-aminobenzotriazole and 4.77g(10mmol) of sodium (6R,7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2-nethoxyininoacetam ido]- 3-acetoxymethyl-ceph-3-em-4-carboxylate were added thereto, and the mixture was stirred at 72 ^* C for 1 hour while adjusting the pH to 6.5— 7.0.

After completion of reaction, the reaction solution was cooled to room teraperature and stirred for 30 min while adjusting its pH to 1—1.5 with 3N hydrochloric acid. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure. The concentrate was subjected to column chromatography over aluminium oxide and silica gel using 80% aqueous acetonitrile as an eluent. The fractions obtained were concentrated under the reduced pressure and dissolved in a small amount of water. This was purified by HP-20(Mitsubishi) column chromatography (eluent: 15% aqueous acetonitrile). The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 1.48g of the desired compound in pale yellowish white amorphous form (28%). Rf = 0.3 (in 80% aqueous acetonitrile) m.p. = 168'C —(decomp.) MS(FAB, M+l) = 530 NMR (DMSO-dβ + 20% DCl, δ)

3.85—4.0 (m, 2H, C-2), 4.05(s, 3H, 0CH ₃ ) 5.05—5.5(q, 2H, C-3), 5.3 (d, IH, C-6)

5.86(d, IH, C-7), 7.04 (s, IH, thiazole-H) 7.65—8.1 (m, 4H, phenyl)

Exarap 1 e 19

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(l-aminobenziraidazol-3-ium) methy1]-ceph-3-em-4-carboxy1ate In a mixed solvent of acetonitrile(lθml) ad water(20ml) was dissolved 16.06g(120mraol) of lithium iodide, the solution was heated to 65 ^* C and 3.2g(24mmol) of 1-arainobenzimidazole and 5.73g(12mmol) of sodium (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacet-am ido]- 3-acetoxyraethyl-ceph-3-em-4-carboxylate were added thereto. The reaction mixture was stirred at 70°C for 1.5 hours while adjusting the pH to 6.5—7.0.

After completion of reaction, the reaction solution was cooled to room temperature and the pH was adjusted to 1—1.5 with 3N hydrochloric acid. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure.

The concentrate was subjected to column chromatography over aluminium oxide and then silica gel (eluent:80% aqueous acetonitrile). The fractions obtained were concentrated under the reduced pressure and dissolved in a small amount of water. This was purified by HP-20 (Mitsubishi) column chromatography using 15% aqueous acetonitrile. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 2.28g of the desired compound in a white amorphous form(36%). Rf = 0.3 (in 80% aqueous acetonitrile) m.p. = 155 ^* C —-(decomp.) MS(FAB, M+l) = 529 NMR (DMS0-d«5, δ)

-3.2—3.7 ( , 2H, C-2), 3.8 (s, 3H, 0CH ₃ ) 5.04 (d, IH, C-6), 5.4— .8(m, 3H, C-3, C-7) 6.75(s, IH, thiazole-H), 7.5—8.5 (ra, 4H, phenyl) 10.15 (s, IH, imidazole)

Example 20

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-methoxyiminoacetaraido]-3-[(l-aminoindazol-2-ium)methyl]- ceph-3-em-4-carboxylate By following the procedure in Example 19 except that 3.2g(24mmol) of 1-arainoindazole was eraployed in place of 1-aminobenzimidazole, there was obtained 1.94g of the desired compound(31%). Rf = 0.31 (in 80X aqueous acetonitrile) NMR (DMSO-dβ + 20% DCl, δ ) 3.87—4.1 (ra, 2H, C-2), 4.02 (s, 3H, OCH3) 5.3 (q, 2H, C-3), 5.33 (d, IH, C-6) 5.82 (d, IH, C-7), 7.03 (s, IH, thiazole-H) 7.1—8.2 (m, 4H, phenyl), 8.8 (s, IH)

Exaraple 21

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-methoxyiminoacetaraido]-3-£(1-amino-l,4,5,6-tetrahydropyr iraldin- 3-ium)methyl]-ceph-3-e_n-4-carboxylate

By following the procedure in Example 19 except that 2.38g(24ramol) of 1-amino-l,4,5,6-tetrahydropyrimidine was eraployed in place of 1-aminobenzimidazole, there was obtained 1.68g of the desired compound (28%).

Rf = 0.29 (in 80% aqueous acetonitrile) NMR (DMS0-d<s + 20% DCl, δ )

1.4—1.8 (q, 2H), 3.3—3.9 (ra, 6H)

3.98 (s, 3H, OCHs), 5.21 (q, 2H, C-3) 5.3 (d, IH, C-6), 5.75 (d, IH, C-7)

7.04 (s, IH, thiazole-H), 8.5 (s, IH)

Preparation Example 1

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- carboxyraethoxyirainoacetamido]-3-acetoxyraethyl-ceph-3-em- 4-carboxylic acid A) In 15ml of N,N-dimethylforraaraide were dissolved 1.09g(2mraol) of (Z)-2-(2-tritylaminothiazol-4-yl)-2-(2-tert-butoxycarbonylme thoxyimino) acetic acid and 0.88g(2mmol) of diphenylmethyl 7 _/ 3-araino-3- acetoxyraethyl-ceph-3-em-4-carboxylate, and the solution was stirred and cooled to 0"C. 0.28g(2.1mraol) of 1-hydroxybenzotriazole and then 0.43g(2. lramol) of dicyclohexylcarbodiiraide -were added thereto. The reaction mixture was stirred at room temperature for 5 hours and left to stand overnight. The reaction mixture was filtered and the filtered solid was washed with diethyl ether. The filtrate and the washing were combined together and 100ml of water added thereto followed by extraction with ethyl acetate. The organic layer was washed with water, IN hydrochloric acid, sodium bicarbonate solution and then saturated brine, separated, dried over anhydrous sodium sulfate and concentrated under the reduced pressure. The concentrate was subjected to silica gel chromatography(eluent: 2.5% methanol/dich1ororaethane). The desired fractions were concentrated under the reduced pressure and

crystallized from diisopropylether to give 1.35g of diphenylmethyl (6R, 7R)-7-[(Z)-2-(2-tritylarainothiazol-4- l)-2-(2-tert-butoxycarbonyl- methoxyimino)acetamido]-3-acetoxynethyl-ceph-3-em-4-carboxyl ate as a desired compound(70%). B) In a mixed solvent of anisole(15ml) and trifluoroacetic acid(15ral) was dissolved 1.35g(1.4mraol) of the product prepared in the above(A) at 0 ^* C and the solution was stirred at room temperature for 2 hours. After completion of the reaction, the reaction solution was concentrated under the reduced pressure and the residue was placed in 150ml of n-hexane cooled to -20'C'—30°C. 70ml of diethyl ether was added thereto and the resulting precipitate was filtered to give 0.75g of (6R, 7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2-carboxymethoxyiminoace taraldo]-3- acetoxynethyl-ceph-3-em-4-carboxylic acid trifluoroacetate. The solid was suspended in 10ml of water, the pH was adjusted to 6.5 with saturated sodium bicarbonate solution and the solution was washed with ethyl acetate. The aqueous layer was adjusted to pH 1.5 and the resulting precipitate was filtered to give 0.48g of (6R, 7R)-7-[(Z)-2-(2-aninothiazol-4-yl)-2-carboxymethoxyiminoacet araido]-3- acetoxyraethyl-ceph-3-em-4-carboxylic acid as a desired compound(68%) . Rf = 0.65 (in 80% aqueous acetonitrile)

2.11 (s, 3H), 3.25—3.82 (q, 2H, C-2)

4.60 (s, 2H), 4.79—4.89 (d, 2H, C-3)

5.19—5.25 (d, IH, C-6), 5.79—5.85 (dd, IH, C-7) 6.71 (s, IH, thiazole-H), 7.27 (br, 2H)

9.47 (d, IH)

Example 22

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-carboxyraethoxyirainoacetamido]-3-[(1-amino-l,2,3,6- tetrahydropyridinium)methyl]-ceph-3-era-4-carboxylate In 15ml of water was dissolved 3.75g(25mmol) of sodium iodide and the solution was heated to 70—-75°C. To this was added 0.59g(6mraol) of 1-amino-l,2,3,6-tetrahydropyridine and 2.5g(5mmol) of (6R,7R)-7-[(Z)- 2-(2-aminothiazol-4-yl)-2-carboxyraethoxyirainoacetamido]-3- acetoxymethyl- ceph-3-em-4-carboxylic acid prepared in Preparation Example 1 was slowly added while adjusting the pH to 6.5—7.0. The mixture was stirred at 70—751C for 2 hours, cooled to room temperature and the pH was adjusted to 1—1.5 with 3N hydrochloric acid. After stirring for 30 rain, the resulting precipitate was filtered off and the filtrate was concentrated under the reduced pressure. A small amount of water was added to the concentrate and chromatography over HP-20(eluent: 80% aqueous acetonitrile) was carried out. The fractions containing desired product were concentrated under - the reduced pressure and freeze-dried to give 0.46g of the desired compound(17%). Rf = 0.28 (in 80% aqueous acetonitrile)

2.1—2.45 (m, 2H), 3.2—4.4 (m, 6H)

4.71 (s, 2H), 4.8—5.2 (m, 2H, C-2) 5.21 (d, IH, C-6), 5.5—5.87 (m, 3H)

6.72 (s, IH, thiazole-H), 7.25 (br, 2H) 9.45 (d, IH, -NH)

Exaraple 23

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-carboxyaethoxyirainoacetamido]-3-(l-aminopiperidiniurameth yl)- cepl.-3-e_π-4-carboxylate By following the procedure in Example 22 except that 0.65ml(6mmol) of l-aminopiperidine was employed in place of 1-araino-l,2,3,6- tetrahydropyridine, there was obtained 0.53g of the desired compound (20%).

Rf = 0.33 (in 80% aqueous acetonitrile) NMR (20% DCl, δ )

1.53—2.22 (m, 6H, piperidine) 3.3—4.5 (a, 6H, C-2, piperidine), 4.7 (s, 2H) 4.88—5.21 (a, 2H, C-3), 5.35 (d, IH, C-6) 5.8 (d, IH, C-7) , 7.2 (s, IH, thiazole-H)

Exaraple 24

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-carboxymethoxyininoacetamido]-3-[(2-araino-l,2,3,4- tetrahydroisoquinolin-2-ium)methy]]-ceph-3-em-4-carboxylate By following the procedure in Example 22 except that 0.89g(6mmol) of 2-amino-l,2,3,4-tetrahydroisoquinoline was employed in place of 1-araino- 1,2,3,6-tetrahydropyridine, there was obtained 0.55g of the desired compound (19%).

Rf = 0.38 (in 80% aqueous acetonitrile) NMR (DMSO-dώ + 20% DCl, δ )

3.1—3.48 (ra, 2H), 3.4—4.4 (ra, 6H) 4.6—5.0 (m, 2H, C-3), 5.2 (d, IH, C-6)

5.75 (d, IH, C-7), 7.05 (s, IH, thiazole-H) 7.12—7.82 (m, 4H, phenyl)

Exaraple 25 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-carboxyraethoxyiminoacetamido]-3-[(l-araino-l-methyl-4- piperazinium)methyl]-ceph-3-era-4-carboxylate

By following the procedure in Example 22 except that 0.91g(6mmol) of l-amino-l-methyl-4-piperazinium chloride hydrochloride was employed in place of 1-amino-l,2,3,6-tetrahydropyridine, there was obtained 1.25g of the desired compound(45%). Rf = 0.11 (in 80% aqueous acetonitrile)

3.0—3.8 (m, 5H), 4.65 (s, 2H) 5.1 (d, IH, C-6), 5.65 (dd, IH, C-7)

6.05 (s, 2H, NH ₂ ), 6.72 (s, IH, thiazole-H) 7.2 (br, 2H), 9.3 (d, IH, -NH)

Exaraple 26 Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-carboxyraethoxyirainoacetamido]-3-[(4-amino-l, ,4-triazo1- 2-ium)methyl]-ceph-3-em-4-carboxylate

In 8ml of water was dissolved 2.25g(15mraol) of sodium iodide and the solution was heated to 70— 5°C. 0.3g(3.6_πraol) of 4-araino-l,2,4- triazole was "added and the pH was adjusted to 6.5—7.0. While maintaining the pH at 6.5—7.0, 1.5g(3mmol) of (6R,7R)-7-[(Z)-2-

(2-aminothiazol-4-yl)-2-carboxyraethoxyirainoacetamido]-3 -acetoxyraethyl-

ceph-3-em-4-carboxylic acid prepared in Preparation Example 1 was slowly added. The reaction solution was stirred at 70—75'C for 2 hours, cooled to room temperature and the pH was adjusted to 1—1.5 with 3N hydrochloric acid. The solution was stirred for 30 rain and the resulting precipitate was filtered off. The filtrate was concentrated, a small amount of water was added and the resulting was subjected to HP-20 chromatography (eluent: 80% aqueous acetonitrile). The fractions containing the desired product were concentrated under the reduced pressure and freeze-dired to give 0.39g of the desired compound (25%). Rf = 0.23 (in 80% aqueous acetonitrile) NMR (DMSO-dύ + 20% DCl, δ )

3.25—3.7 (ra, 2H, C-2), 4.7 (s, 2H)

5.04 (d, IH, C-6), 5.1—5.3 (q, 2H, C-3) 5.65 (d, IH, C-7), 7.02 (s, IH, thiazole-H) 8.75, 9.8 (s.s, 2H, triazole)

Exaraple 27

Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-carboxynethoxyirainoacetamido]-3-[(l-aminoimidazol-3-iuffl ) methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 26 except that 0.3g(3.βramol) of 1-arainoimidazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.31g of the desired compound(20%). Rf = 0.29 (in 80% aqueous acetonitrile) NMR (DMS0-dd + 20% DCl, δ)

3.28—3.8 (m, 2H, C-2), 4.70 (s, 2H)

5.05 (q, 2H, C-3) . 5.22 (d, IH, C-6)

5.75 (d, IH, C-7), 7.0 (s, IH, thiazole-H)

7.6, 8.2 (s,s, 2H, imidazole), 9.0 (s, IH, imidazole)

Example 28 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yi)-

2-carboxyraethoxyirainoacetamido]-3-[(l-arainopyrazol-2-i ura) methy1]-ceph-3-em-4-carboxylate

By following the procedure in Example 26 except that 0.3g(3.6ramol) of 1-aminopyrazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.29g of the desired compound(18%). Rf = 0.29 (in 80% aqueous acetonitrile) NMR (DMSO-dό + 20% DCl, δ )

3.3—3.75 (ra, 2H, C-2), 4.75 (s, 2H) 5.10 (q, 2H, C-3), 5.25 (d, IH, C-6) 5.7 (d, IH, C-7), 6.8 (t, IH, pyrazole)

7.07 (s, IH, thiazol-H), 7.7, 8.5 (s,s, 2H, pyrazole)

Example 29

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-carboxyraethoxyirainoacetamido]-3-[(l-arainoindazol-2-ium) methy1]-ceph-3-em-4-carboxylate

By following the procedure in Example 26 except that 0.48g(3.6ramol) of 1-aminoindazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.32g of the desired compound(19%). Rf = 0.30 (in'80% aqueous acetonitrile) NMR (DMSO-dβ + 20% DCl, δ )

3.85—4.1 (ra, 2H, C-2), 4.76 (s, 2H)

5.31 (q, HI, C-3), 5.4 (d, III, C-6)

5.85 (d, IH, C-7), 7.1 (s, ill, thiazole-H)

7.2—8.18 (ra, 4\i, phenyl) , 8.5 (s, IH, indazole)

Example 30

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-carboxymethoxyininoacetamido]-3-[(l-aminobenzimidazol-3- ium) ethyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 26 except that 0.48g(3.6mmol) of 1-aminobenzimidazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.49g of the desired compound(28%). Rf ~ 0.3 (in 80% aqueous acetonitrile) NMR (DMSO-dβ + 20% DCl, δ )

3.83—4.0 (ra, 2H, C-2), 4.7 (s, 2H) 5.06—5.5 (q, 2H, C-3), 5.19 (d, IH, C-6)

5.79 (d, IH, C-7), 7.01 (s, IH, thiazole-H) 7.5—7.9 (m, 4H, phenyl) , 9.5 (s, IH, imidazole)

Exaraple 31 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-carboxymethoxyirainoacetamido]-3-[(1-amino-l,4,5,6- tetrahydropyrimidin-3-ium)methy]]-ceph-3-em-4-carboxy]ate By following the procedure in Example 26 except that 0.36g(3.6mmol) of 1-amino-l,4,5,6-tetrahydropyrimidine was employed in place of 4- araino-1,2,4-triazole, there was obtained 0.37g of the desired compound (23%). Rf = 0.29 (in 80% aqueous acetonitrile)

NMR -(DMSO-dβ + 20% DCl , δ )

1.5—1. 85 (q, 2H) , 3.3—3. 92 (a, 6H, C-2) 4.69 (s, 2H) , 5.2 (q, 2H, C-3) 5.29 (d, IH, C-6) , 5. 8 (d, IH, C-7) 7.05 (s, IH, thiazole-H) , 7.5 (s, IH)

Preparation Example 2

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetaraldo]-3-acetoxymethyl- ceph-3-em-4-carboxylic acid

A) A solution of 1.14g(2mmol) of (Z)-2-(2-tritylaminothiazol-4-yl)-2- (2-tert-butoxycarbonylprop-2-oxyiraino) acetic acid and 0.88g(2mmol) of 7 β-amino-3-acetoxyraethyl-ceph-3-em-4-carboxylate in 15ml of N,N- dimethylfornaraide was stirred, cooled to 0—5 ^* C, and 0.30g(2.2ramol) of N-hydroxybenzotriazole hydrate and then 0.45g(2.2mmol) of N,N- dicyclohexylcarbodiimide were added thereto. The reaction mixture was stirred at room temperature for 6 hours and- left to stand overnight. The reaction mixture was filtered and the filtered solid was washed with a small amount of diethyl ether. The filtrate and the washing were combined together, lOOral of water was added followed by extraction with ethyl acetate. The organic layer was washed with water, IN hydrochloric acid, sodium bicarbonate solution and then saturated brine, separated, dried over anhydrous sodium sulfate and concentrated under the reduced pressure. The concentrate was subjected to silica gel chromatography (eluent: 2% raethanol/dichloromethane). The fractions containing the desired product were concentrated under the reduced pressure and crystallized from diisopropyl ether to give 1.49g

of diphenylmethyl (6R,7R)-7-[(Z)-2-(2-tritylarainothiazol-4-yl]-2-(2- tert-butoxycarbonylprop-2-oxyimino)acetamido]-3-acetoxyraeth l-ceph-3-em- 4-carboxylate as a desired product(75%).

B) In a mixed solvent of anisole(δml) and trifluoroacetic acid (15ml) was dissolved 1.49g of the product prepared in the above A) at 0°C and the temperature was raised to room temperature. After stirring for 1.5 hours, the reaction solution was concentrated under the reduced pressure and 60ml of diisopropyl ether was added to precipitate 700mg of (6R, 7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2-(2-carboxyprop-2-oxyim ino)acetamido ]-3-acetoxymethyl-ceρh-3-em-4-carboxylic acid trifluoroacetate, which was filtered. Water(2.6ml) was added to the solid and the pH was adjusted to 7.0 with saturated sodium bicarbonate solution. Purification by HP-20 chromatography (eluent: 70% aqueous acetonitrile) gave 0.55g of (6R,7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2-(2-carboxyprop- oxyimino)acetamido]-3-acetoxymethyl-ceph-3-e_π-4- carboxylic acid (70%).

1.32 (d, 6H), 2.22 (s, 3H), 3.52 (q, 2H, C-2) 4.87 (q, 2H, C-3), 5.2 (d, IH, C-6), 5.75 (dd, IH, C-7) 6.87 (s, IH, thiazole-H)

Exaraple 32

Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-[(1-amino-l,2,3,6 - tetrahydropyridiniura)raethyl]-ceph-3-era-4-carboxylate In 15ml of water was dissolved 3.75g(25maol) of sodium iodide and the solution was heated to 70—75°C. 0.59g(6mraol) of 1-amino-l,2,3,6- tetrahydropyridine was added and then, while adjusting the pH to 6.5—

7.0, 2.64g(5mmol) of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-(2- carboxyprop-2-oxyimino)acetamido]-3-acetoxymethyl-ceph-3-em- 4-carboxylic acid prepared in Preparation Example 2 was slowly added. The mixture was stirred at 70— 5°C for 2 hours, cooled to room temperature and the pH was adjusted to 1—1.5 with 3N hydrochloric acid. The solution was stirred for 30 min and the precipitate was filtered off. The filtrate was concentrated under the reduced pressure, a small amount of water was added and subjected to HP-20 chromatography (80% aqueous acetonitrile). The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.51g of the desired compound(18%). NMR (DMSO-dβ + 20% DCl, δ)

1.34 (d, 6H), 2.0—2.4 (m, 2H) 3.1—3.9 (m, 6H, C-2), 4.0—4.3 (ra, IH, C-3) 4.7—5.1 (m, 2H, C-3), 5.2—5.88 (m, 3H, C-7) 6.97 (s, 2H, thiazole-H)

Exaraple 33

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyimino)acetamido]-3-(l-aminopiperidiniu m- methyl)-ceph-3-em-4-carboxylate

By following the procedure in Example 32 except that 0.65ral(6ramol) of 1-aminopiperidine was employed in place of 1-amino-l,2,3,6- tetrahydropyridine, there was obtained 0.53g of the desired compound (19%).

NMR (DMS0-dώ + 20% DCl, δ )

1.30 (d, 6H), 1.5—2.3 (a, 6H)

3.2—4.3 (m, 7H), 4.78—5.1 (ra, IH, C-3) 5.2 (d, IH, C-6), 5.7 (d, IH, C-7) 7.02 (s, IH, thiazole-H)

Example 34

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazoi-4-yl)- 2-(2-carboxyprop-2-oxyimino)acetamido3-3-[(2-amino-l,2,3,4- tetrahydroisoquinolin-2-ium)methyl]-ceph-3-em-4-carboxylate By following the procedure in Exaraple 32 except that 0.89g(6mraol) of 2-amino-l,2,3,4-tetrahydroisoquinoline was employed in place of 1-araino-1,2,3,6-tetrahydropyridine, there was obtained 0.52g of the desired compound(17%). NMR (DMS0-d<s + 20% DCl, δ )

1.28 (d, 6H), 3.1—3.51 (ra, 2H) 3.6—4.3 (m, 7H), 4.5—5.1 (a, IH, C-3) 5.2 (dd, IH, C-6), 5.75 (dd, IH, C-7) 6.95 (s, IH, thiazole-H), 7.1—7.7 (a, 4H, phenyl)

Example 35 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-[(l-amino-l-me thyl- 4-piperaziniura)raethyl]-ceph-3-era-4-carboxylate

By following the procedure in Exaraple 32 except that 0.91g(6mraol) of l-amino-l-methyl-4-piperazinium chloride hydrochloride was employed in place of 1-amino-l,2,3,6-tetrahydropyridine, there was obtained 0.94g of the desired compound(32%) . NMR (DMSO-dβ + 20% DCl, δ )

1.32 (d, 6H), 3.0—3.8 (m, 15H) 4.96 (d, IH, C-6), 5.51 (d, IH, C-7) 7.01 (s, IH, thiazole-H)

Example 36

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyimino)acetamido]-3-[(4-amino-l,2,4- triazol-2-ium)methyl]-ceph-3-em-4-carboxylate In 8ml of water was dissolved 2.75g(15ramol) of sodium iodide, the solution was heated to 70—75T. and 0.3g(3.6mmol) of 4-amino-l,2,4- triazole was added. While adjusting the pH to 6.5—7, 1.5g(3mmol) of (6R,7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2-(2-carboxyprop-2-o xyimino) acetaraido]-3-acetoxymethyl-ceph-3-era-4-carboxylic acid prepared in Preparation Example 2 was slowly added. The mixture was stirred at 70- —75 ^* C for 3 hours, cooled to room temperature and the pH was adjusted to 1—-1.5 with 3N hydrochloric acid. The mixture was stirred for 30 min and the resulting precipitate was filtered off. The filtrate was concentrated under the reduced pressure, a small amount of water was added and subjected to HP-20 chromatography (80% aqueous acetionitrile). The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.35g of the desired compound(20%). MS(FAB, M+l) = 552 NMR (DMSO-dή + 20% DCl, δ ) 1.30 (d, 6H), 3.3—3.6 (m, 2H, C-2)

5.07 (d, IH, C-6), 5.0—5.37(q, 2H, C-3) 5.7 (d, IH, C-7) , 7.01 (s, IH, thiazole-H)

8.7, 9.8 (s.s, 211, triazole)

Example 37

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-[(l-aminoinidazol- 3-1um)methy1]-ceph-3-em-4-carbox 1ate

By following the procedure in Example 36 except that 0.3g(3.6ramoI) of 1-arainoimidazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.36g of the desired compound(23%). NMR (DMSO-d + 20% DCl, δ )

1.33 (d, 6H), 3.3—3.7 (m, 2H, C-2) 4.9—5.3 (m, 3H, C-6, C-3), 5.7 (d, IH, C-7) 6.98 (s, IH, thiazole-H), 7.7, 8.6 (s,s, 2H) 9.1 (t, IH)

Example 38

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothi-azol-4-y] - 2-(2-carbo yprop-2-ox iaino acetaraido]-3-_. l-aminopy^azol- 2-ium)methyl]-ceph-3-em-4-carboxylate By following the procedure in Example 36 except that 0.3g(3.6ramol) of 1-aminopyrazole was eraployed in place of 4-amino-l,2,4-triazole, there was obtained 0.34g of the desired compound(22%). NMR (DMSO-de + 20% DCl, δ)

1.3 (d, 6H), 3.2—3.7 (m, 2H, C-2) 4.9—5.25 (ra, 3H, C-3, C-6), 5.65 (d, IH, C-7) 6.75 (s, IH), 7.01 (s, IH, thiazole-H) 7.8, 8.5 (s,s, 2H)

Example 39

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-[(l-aminoindaz ol-

2-ium)methy1]-ceph-3-em-4-carboxy1ate By following the procedure in Example 36 except that 0.48g(3.δramol) of 1-aminoindazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.35g of the desired compound(20%). NMR (DMSO-dό + 20% DCl, δ )

1.32 (d, 6H), 3.87—4.1 (ra, 2H, C-2) 5.1—5.4 (m, 3H, C-3, C-6), 5.80 (d, IH, C-7)

7.03 (s, IH, thiazole-H), 7.1—8.2 (a, 4H, phenyl)

8.9 (s, IH)

Exaraple 40 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-(2-carboxyprop-2-oxyiraino)acetamido]-3-[(l-aminobenzir aidzol- 3-ium)methy1]-ceph-3-em-4-carboxylate

By following the procedure in Example 36 except that 0.48g(3.6ramol) of 1-aminobenziraidazole was eraployed in place of 4-amino-l,2,4-triazole, there was obtained 0.3δg of the desired compound(22%). NMR (DMSO-dβ + 20% DCl, δ )

1.29 (d, 611), 3.2—3.8 (m, 211, C-2) 5.1—5.4 (m, 3H, C-3, C-6), 5.7 (d, IH, C-7) 7.03 (s, IH, thiazole-H), 7.35—8.01 (a, 4H, phenyl) 9.7 (s, IH,' iraidazole-H)

Exaraple 41

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-[(l-amino-l,4,5,6- tetrahydropyrimidin-3-ium) ethyl]-ceph-3-em-4-carboxylate By following the procedure in Example 36 except that 0.36g(3.6ramol) of 1-amino-l,4,5,6-tetrahydropyrimidine was eraployed in place of

4-amino-l,2,4-triazole, there was obtained 0.31g of the desired compound(19%).

NMR (DMSO-dβ + 20% DCl, δ ) 1.32 (d, 6H), 1.4—1.8 (q, 2H)

3.3—3.9 (m, 6H, C-2), 5.15—5.4 ( , 3H, C-3, C-6) 5.76 (d, IH, C-7), 7.01 (s, IH, thiazole-H) 8.5 (s, IH)

Preparation Example 3

Synthesis of (6R,7R)-7-[(Z)-2-(2-ami othiazol-4-yl)- 2-(2-carboxypro -2-oxyimino)acetamido]-3-acetox methyl- ceph-3-em-4-carboxylie acid A) In 15ml of N,N-diraethylformamide was dissolved 1.22g(3mmol) of (Z)-2-[(2-chloroacetamidothiazol-4-yl)-2-(2-tert-butoxycarbo nylprop-2- oxyimino)acetic acid, 0.66g(3.21maol) of N,N-dicyclohexylcarbodiimide was added thereto and the aixture was stirred at room temperature for 2 hours. 1.41g(3.21ramol) of diphenylmethyl 7 _/ 3-araino-3-acetσxyraethyl- ceph-3-em-4-carboxylate was added thereto at 0—5 ^* C and the mixture was stirred at the same temperature for 4.5 hours.

After completion of the reaction, the insoluble was filtered off, 200ral of dichlororaethane was added to the filtrate and this was washed

6δ

with diluted hydrochloric acid and then with water. The organic layer was dried over anhydrous magnesium sulfate and concentrated under the reduced pressure. The residue was purified by silica gel column chromatography using 6% methanol/dichloromethane as an eluent to give l.δ6g of diphenylmethyl (6R,7R)-7-[(Z)-2-(2-chloroacetamidothiazol-4- yl)-2-(2-tert-butoxycarbony1prop-2-oxyiraino)acetamido]-3-ac etoxymethyl- ceph-3-em-4-carboxylate in pale yellow powder forra(70%). B) In 15ml of anisole was dissolved the product obtained in the above A), the solution was cooled to 0—5°C and 15ral of trifluoroacetic acid was added thereto. The reaction mixture was stirred at the same temperature for 2 hours.

After completion of the reaction, the reaction solution was concentrated under the reduced pressure to remove trifluoroacetic acid.

The residue was cooled to -20——10°C, 200ml of diisopropyl ether was added and the mixture was stirred at the same temperature for 1 hour.

The resulting precipitate was filtered, washed thoroughly with diisopropyl ether and dried to give 1.37g of (6R,7R)-7-[(Z)-2-(2- chloroacetamidothiazol-4-yl)-2-(2-carboxyprop-2-oxyimino)ace tamido]-3-ac etoxymethyl-ceph-3-em-4-carboxylic acid trifluoroacetate (δ5%). C) The product obtained in the above B) was suspended in 70ml of water and the pH was adjusted to 6.5—7.0 with saturated sodium bicarbonate solution. 0.26g(2mmol) of sodium N-methyl-dithiocarbamate was added thereto and the mixture was stirred at room temperature for 5 hours. After completion of the reaction, the reaction mixture was washed with ethyl acetate and the aqueous layer containing the desired product was concentrated under the reduced pressure. The concentrate was

purified by HP-20 column chromatography. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.73g of (6R,7R)-[(Z)-2-(2-aminothiazol-4-yl)-2- (2-carboxyprop-2-oxyinino)acetanido]-3-acetoxymethy]-ceph-3- em-4- carboxylic acid(72%).

Rf = 0.5 (in δ0% aqueous acetonitrile)

The NMR spectra of the desired compound were the same as those of the compound obtained in Preparation Example 2.

Example 42

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-[(4-aminomorpholi n- 4-ium) ethyl]-ceph-3-em-4-carboxylate

In a mixed solvent of acetonitrile(lθral) and water(lθml) was dissolved 6.69g(50mmol) of lithium iodide, 1.02g(10amol) of 4-amino morpholine was added thereto and the mixture was heated to 65—βδ'C Water ral) containing 2.64g(5maol) of (6R,7R)-7-[(Z)-2-(2-aminothiazol- 4-yl)-2-(2-carboxyprop-2-oxyimino)acetaraido]-3-acetoxymethy l-ceph-3-en-4 -carboxylic acid prepared in Preparation Exaraple 3 and 0. lg of sodium hydroxide was added thereto and the reaction mixture was stirred at 70— 72 ^* C for 1.5 hours while adjusting the pH to 6.5—7.0.

After completion of the reaction, the reaction solution was cooled to room temperature and the pH was adjusted to 1.5 with 3N hydrochloric acid. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure followed by purification by column chromatography over aluminium oxide (eluent: 80% aqueous acetonitrile). The fractions were concentrated under the reduced pressure, dissolved in

a small amount of water and purified by HP-20 column chromatography. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.47g of the desired compound (16%). NMR (DMSO-dύ + 20% DCl, δ)

1.3 (d, 6H), 3.35—4.55 (ra, 11H)

4.95—5.36 (q, IH, C-3), 5.34 (d, IH, C-6)

5.8 (d, IH, C-7), 7.02 (s, IH, thiazole-H)

Exaraple 43

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-(l- aminopyrrolidiniuramethyl)-ceph-3-em-4-carboxylate

By following the procedure in Example 42 except that 1.23g(10mmol) of 1-aminopyrrolidine was employed in place of 4-aminomorpholine, there was obtained 0.51g of the desired compound(18%). NMR (DMSO-dβ + 20% DCl, δ )

1.34 (d, 6H), 1.75—2.25 (a, 4H) 2.92—3.75 (a, 6H) , 3.δ—4.2 (a, IH, C-3) 4.4—5.0 (m, IH, C-3), 5.2 (d, IH, C-6)

5.7 (d, IH, C-7) _r 7.02 (s, IH, thiazole-H)

Exaraple 44

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carb ^' oxyprop-2-oxyiraino)acetaraido]-3-[(l-amino-3- pyrrolin-l-iura)raethyl]-ceph-3-em-4-carboxylate By following the procedure in Example 42 except that 0.δ4g(10ramol)

of l-amino-3-pyrroline was employed in place of 4-arainomorpholine, there was obtained 0.39g of the desired compound(14%). NMR (DMSO-da + 20% DC], δ )

1.30 (d, 6H), 3.6—4.1 (m, 3H, C-2, C-3) 4.65 (s, 4H), 4.57—5.1 (a, IH, C-3)

5.2 (d, IH, C-6), 5.65 (d, IH, C-7)

7.03 (s, IH, thiazole-H)

Exaraple 45 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-y])- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-(l- aminoindoliniummethyl)-ceph-3-era-4-carboxylate

By following the procedure in Example 42 except that 1.34g(10mmol) of 1-aminoindoline was employed in place of 4-arainoraorpholine, there was obtained 0.4δg of the desired compound(16%). NMR (DMSO-dβ ^• + 20% DCl, δ )

1.32 (d, 6H), 2.δ—3.2 (a, 2H) 3.4—4.5 (m, 5H), 4.6—5.2 (m, 2H, C-3, C-6) 5.65 (d, IH, C-7), 7.01 (s, IH, thiazole-H) 7.05—7.δ (ra, 4H, phenyl)

Example 46

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-(l- aminopyrroliuramethyl)-ceph-3-em-4-carboxylate

By following the procedure in Example 42 except that 0.δ2g(10ramol) of 1-aminopyrrole was employed in place of 4-aminomorpholine, there was

obtained 0.45g of the desired compound(16%) . NMR (DMS0-d ₆ + 20% DCl, )

1.28 (d, 6H), 3.64 (a, 3H, C-2, C-3)

4.6—5.2 (a, 2H, C-3, C-6), 5.6 (d, IH, C-7) 6.2 (t, 2H), 6.6 (t, 2H)

7.03 (s, IH, thiazole-H)

Example 47

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-(l- arainoindoliumraethyl)-ceph-3-em-4-carboxylate

By following the procedure in Example 42 except that 1.32g(10mmol) of 1-aminoindole was employed in place of 4-arainomorpholine, there was obtained 0.49g of the desired compound(16%). NMR (DMS0-dύ + 20% DCl, δ)

1.29 (d, 6H), 3.5—4.0 (m, 3H, C-2, C-3) 4.6—5.2 (m, 2H, C-3, C-6), 5.65 (d, IH,- C-7) 6.5 (d, IH), 7.02 (s, IH, thiazole-H) 7.05—7.9 (ra, 5H, phenyl)

Example 48

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyimino)acetaraido]-3-[(l-amino-4- hydroxypiperidinium)methyl]-ceph-3-em-4-carboxylate By following the procedure in Example 42 except that 1.16g(10ramol) of l-amino-4-hydroxypiperidine was eaployed in place of 4- arainomorpholine, there was obtained 0.51g of the desired compound(17%) .

Rf = 0.14 (in 80% aqueous acetonitrile) NMR (20% DCl, δ )

1.18—2.0 (m, 4H), 1.30 (d, 6H)

3.2—4.3 (a, 7H), 4.86—5.2 (m, 2H, C-3) 5.35 (d, IH, C-6), 5.93 (d, IH, C-7)

7.21 (s, IH, thiazole-H)

Example 49

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-[(1-amino-l,2,3- triazo1-3-ium)methy1]-ceph-3-em-4-carboxy1ate

In a mixed solvent of acetonitrile(lθral) and water(15ml) was dissolved 5.35g(40mmol) of lithium iodide, 0.673g(8ramol) of 1-amino- 1,2,3-triazole was added thereto and the mixture was heated to 65—δδ'C. 2.11g(4mmol) of (6R,7R)-7-[(Z)-2-(2-arainothiazol-4-yl)-2-(2-carboxy- prop-2-oxyiraino)acetaraido]-3-acetoxymethyl-ceph-3-em-4-car boxylic acid prepared in Preparation Example 3 was added -and the mixture was stirred at 70—72"C for 2 hours while adjusting the pH to 6.5—7.0.

After completion of the reaction, the reaction solution was cooled to room temperature and the pH was adjusted to 1—1.5 with 3N hydrochloric acid. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure followed by column chromatography over aluminium oxide using δ0% aqueous acetonitrile solution as an eluent. The fractions were concentrated under the reduced pressure, dissolved in a small amount of water and purified by HP-20 column chromatography. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to

give 0.68g of the desired compound(31%). NMR (DMSO-da + 20% DCl, δ )

1.3 (d, 6H), 3.4—3.7 (m, 2H, C-2)

5.05—5.18 (m, 3H, C-3, C-6), 5.67 (d, IH, C-7) 7.02 (s, IH, thiazole-H), 7.8, 8.7 (s,s, 2H, triazole-H)

Exaraple 50

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-[(l-aminobenzotri azol- 3-ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Exaraple 49 except that 1.07g(8mmol) of 1-arainobenzotriazole was eraployed in place of 1-amino-l,2,3-triazole, there was obtained 0.53g of the desired compound(22%). MS(FAB, M+l) = 602 NMR (DMSO-dώ + 20% DCl. δ )

1.31 (d, 6H), 3.86—4.1 (ra, 2H, C-2) 5.05—5.55 (m, 3H, C-3, C-6), 5.δ7 (d, IH, C-7) 7.02 (s, IH), 7.7—8.15 (ra, 4H, phenyl)

Exaraple 51

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetaraido]-3-[(1-amino-l,2,4- triazol-4-ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 49 except that 0.67g(8mraol) of 1-amino-l,2,4-triazole was employed in place of 1-amino-l,2,3-triazole, there was obtained 0.5g of the desired compound(23%) . NMR (DMSO-dώ + 20% DCl, )

1.28 (d, 6H), 3.29—3.74 (a, 2H, C-2) 5.05—5.25 (m, 3H, C-3, C-6), 5.7 (d, IH, C-7)

6.94 (s, IH, thiazole-H), 8.74, 9.68 (s,s, 2H, triazole)

Exaraple 52

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-(2-carboxyprop-2-oxyiraino)acetamido]-3-[(l-aminotetrazo]- 4- ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 49 except that 0.69g (βmmol) of 1-aminotetrazole was employed in place of 1-aaino-l,2,3-triazole, there was obtained 0.4δg of the desired coapound (22%). NMR(DMS0-d6 + 20% DCl, δ )

1.29 (d, 3H), 3.3—3.76 (ra, 2H, C-2) 5.05—5.2 (ra, 3H, C-3, C-6), 5.7 (d, IH, C-7) 6. 99 (s, IH, thiazole-H) , 9.52 (s, IH, tetrazole-H)

Preparation Exaraple 4

Synthesis of (6R,7R)-7-[(Z)-2-(2-aainothiazoI-4-yl)- 2-carboxyraethoxyirainoacetaaido]-3-acetoxyraethyl- ceph-3-em-4-carboxy]ic acid

A) In 15ml of N,N-dinethylformamide was dissolved 1.13g(3mmol) of (Z)-2-[(2-chloroacetamidothiazol-4-yl)-2-tert-butoxycarbonyl methoxy- imino]acetic acid, and 0.43g(3. lβmmol) of N-hydroxybenzotriazole and 0.66g(3.21mraol) of N,N-dicyclohexy1carbodiimide were added thereto. The reaction mixture was stirred at room teaperature for 2 hours.

1.41g(3.21mmol) of diphenylraethyl 7 _/ 5-amino-3-acetoxyaethyl-ceph-3- era-4-carboxylate was added at 0—-5°C and the aixture was stirred at the

same temperature for 5 hours. After completion of the reaction, the procedure in Preparation Example 3-A) was repeated to give 1.67g of diphenylmethyl(6R,7R)-7-[(Z)-2-(2-chloroacetamidothiazol-4-y l)-2-tert- butoxycarbonylmethoxyiminoacetaraido]-3-acetoxymethyl-ceph-3 -era-4- carboxylate(65%) .

B) The product obtained in the above A) was dissolved in 120ml of anisole and the mixture was cooled to 0—5°C. 12ml of trifluoroacetic acid was added and the reaction mixture was stirred at the same temperature for 2 hours. After completion of the reaction, the procedure in Preparation Example 3-B) was repeated to give 1.2g of (6R, 7R)-7-[(Z)-2-(2-chloroacetaraidothiazol-4- l)-2-carboxymethoxyiminoacetara ido]-3-acetoxymethyl-ceph-3-era-4-carboxylic acid trifluoroacetate(63%) .

C) The product obtained in the above B) was suspended in 65ral of water and the pH was adjusted to 6.5—7.0 with saturated sodium bicarbonate solution. 0.34g(2.6mmol) of sodium N-methyl-dithiocarba- mate was added and the reaction mixture was stirred at room temperature for 4—5 hours. After completion of the reaction, the procedure in Preparation Exaraple 3-C) was repeated to give 582mg of (6R,7R)-7-[(Z)- 2-(2-aminothiazol-4-yl)-2-carboxyraethoxyirainoacetamido]-3- acetoxyraethyl- ceph-3-em-4-carboxylic acid (67%).

Rf = 0.65 (in δ0% aqueous acetonitrile)

2.11 (s, 3H), 3.25-3.62 (q, 2H, C-2) 4.6 (s, 2H), 4.79-4.69 (d, 2H) 5.19—5.2.5 (d, IH, C-6), 5.79—5.85 (dd, IH, C-7) 6.71 (s, IH, thiazole-H), 7.27 (br, 2H, NH ₂ ) 9.5 (d, IH, -NH)

Exaraple 53

Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-carboxynethoxyirainoacetamido]-3-[(4-aminomorpholin-

4-ium)methyl]-ceph-3-em-4-carboxylate In a mixed solvent of acetonitrile(9ml) and water(10ml) was dissolved 6.69g(50mmol) of lithium iodide, the solution was heated to 65—68 ^* C and 1.02g(10mmol) of 4-aminomorpholine was added thereto. Water (3ml) containing 2.5g(5mraol) of (6R,7R)-7-[(Z)-2-(2-aminothiazol- 4-yl)-2-carboxyraethoxyiminoacetamido]-3-acetoxynethyl-ceph- 3-em-4- carboxylic acid and O.lg of sodiura hydroxide was added and the reaction mixture was stirred at 70—73^ for 2 hours while the pH was adjusted to 6.5—7.0.

After completion of the reaction, the reaction solution was cooled to room temperature and the pH was adjusted to 1.5 with 3N hydrochloric acid. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure followed by aluminium oxide column chromatography using δ0% aqueous acetonitrile as an eluent. The fractions were concentrated under the reduced pressure, dissolved in a small amount of water and purified by HP-20 column chromatography. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.79g of the desired compound(29%) .

Rf = 0.34 (in δ0% aqueous acetonitrile) NMR (20% DCl, δ ) 3.4—4.6 (m, 11H, morpholine, C-2, C-3), 4.7 (s, 2H)

5.0—5.35 (q, IH, C-3), 5.33 (d, IH, C-6)

5.8 (d, IH, C-7), 7.18 (s, IH, thiazole-H)

Example 54

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-carboxymethoxyiminoacetamido]-3-(l-aminopyrrolidiniumme thyl)- ceph-3-em-4-carboxylate By following the procedure in Example 53 except that 1.23g(10ramol) of 1-aminopyrrolidine was eraployed in place of 4-aminomorpholine, there was obtained 0.49g of the desired compound(19%). Rf = 0.3 (in 80% aqueous acetonitrile) NMR (20% DCl, δ ) 1.75—2.3 (ra, 4H, pyrrolidine) , 2.91—3.8 (ra, 6H, C-2, pyrrolidine)

3.9—4.4 (m, IH, C-3), 4.65 (s, 2H)

4.5—4.95 (m, IH, C-3), 5.15 (dd, IH, C-6)

5.7 (dd, IH, C-7), 7.12 (s, IH, thiazole-H)

Example 55

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazoi-4-yl)- 2-carboxymethoxyirainoacetamido]-3-[(l-ara-ino-3-pyrrolin-l- ium)methy1]-ceph-3-em-4-carboxylate

By following the. procedure in Example 53 except that 0.84g(10mmol) of l-amino-3-pyrroline was employed in place of 4-arainomorpholine, there was obtained 0.47g of the desired compound(lδ%). Rf = 0.3 (in 80% aqueous acetonitrile) NMR (DMSO-dβ + 20% DCl, δ )

3.5—4.1 (m, 3H, C-2, C-3), 4.6 (s, 4H, pyrroline) 4.56—5.1 ^' (m, IH, C-3), 4.7 (s, 2H) 5.16 (d, IH, C-6), 5.7 (d, IH, C-7) 5.92 (s, 2H, pyrroline), 7.04 (s, IH, thiazole-H)

Exaraple 56

Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazoi-4-yl)-

2-carboxynethoxyiminoacetaaido]-3-(l-aainoindoliniuraaeth yl)-

-ceph-3-era-4-carboxy1ate By following the procedure in Example 53 except that 1.34g(10ramol) of 1-aminoindoline was employed in place of 4-aminomorρholine, there was obtained 0.5g of the desired corapound(17%) . Rf = 0.37 (in 80% aqueous acetonitrile) NMR (DMSO-dc + 20% DCl, δ) 2.8—3.2 (m, 2H, indole)

3.4—4.4 (m, 5H, indole, C-2, C-3), 4.65 (s, 2H)

4.7—5.1 (m, IH, C-3), 5.12 (d, IH, C-6)

5.7 (d, IH, C-7), 7.02 (s, IH, thiazole-H)

7.04-7.6 (m, 4H, phenyl)

Exaraple 57

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothLazol-4-yl)-

2-carboxyraethoxyirainoacetamido]-3-(l-aainopyrroliumraet hyl)-

-ceph-3-era-4-carbox late By following the procedure in Example 53 except that 0.δ2g(10ramol) of 1-aminopyrrole was employed in place of 4-aminomorpholine, there was obtained 0.43g of the desired compound(16%). Rf = 0.2δ (in 80% aqueous acetonitrile) NMR (DMSO-dώ + 20% DCl, δ) 3.6—4.0 (m, 3H, C-2, C-3) , 4.65 (ε, 2H)

4.6—5.0 (a, IH, C-3), 5.2 (d, IH, C-6)

5.65 (d, IH, C-7), 6.12 (t, 2H, pyrrole)

6.5 (t, 2H, pyrrole), 7.03 (s, IH, thiazole-H)

Example 58

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-carboxymethoxyirainoacetamido]-3-(l-aminoindoliummethyl)- -ceph-3-era-4-carboxy1ate

By following the procedure in Example 53 except that 1.32g(10ramol) of 1-aminoindole was employed in place of 4-aminomorpholine, there was obtained 0.41g of the desired compound(14%). Rf = 0.35 (in 80% aqueous acetonitrile) NMR (DMSO-dώ + 20% DCl, δ )

3.47—4.0 (m, 3H, C-2, C-3), 4.62—5.0 (m, IH, C-3) 4.7 (s, 2H), 5.2 (d, IH, C-6) 5.55 (d, IH, C-7), 6.47 (d, IH, indole) 7.01 (s, IH, thiazole-H), 7.03—8.0 (ra, 5H, phenyl)

Example 59

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-carboxyraethoxyirainoacetamido] -3-1(l-araino-4- hydroxypiperidinium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 53 except that 1.16g(10ramol) of l-amino-4-hydroxypiperidine was employed in place of 4- arainoraorpholine, there was obtained 0.56g of the desired compound(20%) . Rf = 0.14 (in 80% aqueous acetonitrile) NMR (20% DC], δ )

1.2—2.0 (m, 4H), 3.22—4.4 (m, 7H)

4.70 (s, 2H), 4.85—5.21 (m, 2H, C-3)

5.35 (d, IH, C-6), 5.95 (d, IH, C-7) 7.22 (s, IH, thiazole-H)

Example 60 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-carboxymethoxyiminoacetamido]-3-[(1-amino-l,2,3- triazol-3-1um)methyl]-ceph-3-em-4-carboxylate

In a mixed solvent of acetonitrile (9ml) and water(15ml) was dissolved 5.35g(40mmol) of lithium iodide, 0.673g(8mmol) of 1-amino- 1,2,3-triazole was added thereto and the mixture was heated to 68 ^C C. 2g(4mraol) of (6R,7R)-7-[(Z)-2-(2-arainothiazoJ-4-yl)-2-carboxymethoxy- irainoacetamido]-3-acetoxyraethyl-ceph-3-em-4-carboxylic acid prepared in Preparation Exaraple 4 was added and the mixture was stirred at 70—72°C for 1.5 hours while adjusting the pH to 6.5—-7.0. After completion of the reaction, the reaction solution was cooled to room temperature and the pH was adjusted to 1.5 with 3N hydrochloric acid. The insoluble was filtered off and tlie filtrate was concentrated under the reduced pressure followed by aluminium oxide column chromatography using 80% aqueous acetonitrile as an eluent. The fractions were concentrated under the reduced pressure, dissolved in a small amount of water and purified by HP-20 column chromatography. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.48g of the desired compound(23%). Rf = 0.25 (in ^' 80% aqueous acetonitrile) NMR (DMSO-dc + 20% DC], δ )

3.4—3.72 (ra, 2H, C-2), 4.75 (s, 2H) , 5.13 (q, 2H, C-3)

δ2

5. 16 (d, III,- C-6) , 5. 7 (d, HI, C-7)

7.02 (s, IH, thiazole-H) , 7. δ , δ . δ ( s, s, 2H, triazol e)

Example 61 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-carboxyraethoxyirainoacetaaido]-3-[(l-arainobenzotriazo l- 3-ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 60 except that 1.07g(δmmol) of 1-arainobenzotriazole was employed in place of 1-amino-l,2,3-triazole, there was obtained 0.51g of the desired compound(22%). Rf = 0.3 (in δ0% aqueous acetonitrile) MS(FAB, M+l) = 574 NMR (DMSO-dύ + 20% DCl, δ)

3.δ2—4.1 (m, 2H, C-2), 4.72 (s, 2H) 5.1—5.45 (q, 2H, C-3), 5.35 (d, IH, C-6)

5.δ5 (d, IH, C-7), 7.02 (s, IH, thiazole-H) 7.5—δ.2 (m, 4H, phenyl)

Example.62 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-carboxyraethoxyirainoacetamido]-3-[(l-amino-l,2,4- triazol-4-ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 60 except that O.67g(βmmol) of 1-amino-l,2,4-triazole was employed in place of 1-amino-l,2,3-triazole, there was obtained 0.53g of the desired compound(25%) . Rf = 0.3 (in 60% aqueous acetonitrile) NMR (DMS0-dώ + 20% DCl, )

3.4—3.8 (a,- 211, C-2), 4.65 (s, 211) 5.12 (q, 2H, C-3) , 5.2 (d, IH, C-6)

5.6 (d, IH, C-7), 6.99 (s, IH, thiazole-H)

8.75, 9.68 (s.s, 211, triazole)

Example 63

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yJ)- 2-carboxyraethoxyiminoacetamido]-3-[(l-arainotetrazol-4- ium)methyl]-ceph-3-em-4-carboxyl te By following the procedure in Exaraple 60 except that 0.69g(8mmo]) of 1-aminotetrazole was-employed in place of 1-araino-l, ,3-triazole, there was obtained 0.43g of the desired compound(19%). Rf = 0.24 (in δ0% aqueous acetonitrile) NMR (DMSO-dβ + 20% DC], δ ) 3.23—3.δ (ra, 2H, C-2), 4.6δ (s, 2H) 5.1 (q, 2H, C-3), 5.25 (d, IH, C-6)

5.7 (d, IH, C-7), 7.05 (s, IH, thiazole-H) 9.6 (s, IH, tetrazole-H)

Preparation Exaraple 5

Synthesis of (6R,7R)-7-[(Z)-2-(2-tritylaminothiazol- 4-yl)-2-methoxyiminoacetaraido]-3-chloromethyl-ceph- 3-era-4-carboxy1ate

To lOral of dichlororaethane were added 0.45g(lramol) of diphenylaethyl 7/9-amino-3-chloromethyl-ceph-3-era-4-carboxylate hydrochloride and 0.53g

(1.2mraol) of (Z)-2-(2-tritylaminothiazol-4-yl)-2-methoxyiminoacetic acid. 0.4ral(5mmol) of pyridine and 0.11ml (1.2mmol) of phosphorus

δ4

oxychloride were slowly added under ice-cooling and the mixture was stirred for 15min. 50ml of chloroform was added, and the mixture was washed with water and dried over anhydrous sodium sulfate.

The solution was concentrated under the reduced presure and subjected to silica gel chromatography (2% methanol/dichlororaethane). The fractions containing the desired product were concenbralcd under the reduced pressure and crystallized from diisopropyl ether to give 0.6g of the desired compound(72%) . NMR (CDCls, δ ) 3.55 (bs, 2H), 4.05 (s, 3H) , 4.30 (bs, 2H) 5.05 (d, IH), 5.90 (dd, IH), 6.75 (s, IH) 6.95 (s, IH), 7.1—7.δ (m, 26H)

Preparation Exaraple 6 Synthesis of diphenylmethyl (6R,7R)-7-[(Z)-2-(2- tritylaminothiazol-4-yl)-2-raethoxyirainoacetamido]- 3-ch1oromethy1-ceph-3-em-4-carbox 1ate -

To 25ml of N.N-diraethylformamide were added 1.3g(3mraol) of (Z)-2-(2- tritylarainothiazol-4-yi)-2-raethoxyirainoacetic acid and 1.35(3mmol) of diphenylmethyl 7 β-amino-3-chlororaethyl-ceph-3-em-4-carboxylate hydroch¬ loride. The mixture was cooled to 0°C, and 0.53g(3.9ramol) of N-hydroxybenzotriazole hydrate and then O.δOg (3.9mraol) of N,N-dicyclo- hexy1carbodiimide were added thereto.

The reaction mixture was stirred for 5 hours and left to ¹ stand overnight. ^" The reaction mixture was filtered and the filtered solid was washed with a small amount of diethyl ether. The filtrate and the washing were combined together and 125ml of water was added thereto.

The reaction mixture was extracted with ethyl acetate and washed with sodium bicarbonate solution and then saturated brine. The organic layer was dried over anhydrous sodium sulfate, concentrated under the reduced pressure and subjected to silica gel chromatography (2% methanol/ dlchloromethane). The fractions containing the desired product were concentrated under the reduced pressure and crystallized from diisopropyl ether to give 1.92g of the desired compound(76%). NMR (CDCls, δ )

3.55 (bs, 2H), 4.05 (s, 3H), 4.30 (bs, 2H) 5.05 (d, IH), 5.90 (dd, IH), 6.75 (s, IH)

6.95 (s, IH), 7.1—7.6 (m, 26H)

Example 64

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-methoxyiminoacetamido]-3-(l-aminopiperidiniuaraethyl)- ceph-3-em-4-carboxyl te A) In 5ml of N,N-dimethylforraaraide were dissolved 2.52g(3.0mmol) of diphenylmethyl (6R,7R)-7-[(Z)-2-(2-tritylaminothiazol-4-yl)-2-methoxy- irainoacetamido]-3-chloromethyl-ceph-3-era-4-carbox late prepared in Preparation Example 5 or 6 and 0.67g(0.45mmol) of sodiura iodide and 0.39ml (3.6mmol) of 1-aminopiperidine was added ^' thereto. The mixture was stirred at room temperature for 3 hours. To the reaction solution was added 15ml of water and the mixture was extracted with dlchloromethane. The organic layer was washed with water, dried ' over anhydrous magnesium sulfate, concentrated under the reduced pressure and subjected to silica gel chroaatography (4% aethanol/dichlororaethane) . The fractions containing the desired product were concentrated under the

66

reduced pressure to give 1.86g of diphenylmethyl (6R,7R)-7-[(Z)-2-(2- tritylaminothiazol-4 yl)-2-raethoxyiminoacetamido]-3-(l-aminopiperidinium methyl)-ceph-3-em-4-carboxylate iodide(60%).

B) To 1.7ml of anisole was added 0.6g(0.5δmmol) of the product obtained in the above A), 5.6rai of trifluoroacetic acid was slowly added under ice-cooling and the mixture was stirred for 1 hour. Diisopropyl ether was added, and the resulting precipitate was filtered and dried. A small amount of water was added to the solid, and the pH was adjusted to 5—8 with saturated sodiura bicarbonate solution. HP-20 colurn chromatography was carried out by using δ0% aqueous acetonitrile solution as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.24g of the desired compound (δ2%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 3.

Exaraple 65

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yD- 2-methoxyiminoacetaraido]-3-[(4-aminomorpholin-4-ium) methy1]-ceph-3-em-4-carboxylate

By following the procedure in Example 64 except that 0.35g(3.6ramol) of 4-arainoraorpholine was employed in place of 1-aminopiperidine, there was obtained 0.63g of the desired compound (43%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 2.

δ7

Example 66

Synthesis of (6R,7R)-7-[(Z)-2-(2-aainothiazol-4-yl)- 2-methoxyiminoacetamido]-3-(l-aminopyrrolidiniumaethyl)- ceph-3-em-4-carboxylate By following the procedure in Example 64 except that 0.44g(3.6mmol) of 1-aminopyrrolidine was employed in place of 1-aminopiperidine, there was obtained 0.65g of the desired compound (45%).

The NMR spectra of the desired compound were the sane as those of the compound prepared in Example 1. Exaraple 67

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazoi-4-yl)- 2-methoxyiminoacetamido]-3-[(1-amino-l,2,3,6-tetrahydro- pyridiniun)raethyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 64 except that 0.53g(3.6mmol) of 1-amino-l,2,3,6-tetrahydropyridine was employed in place of 1- aminopiperidine, there was obtained 0.54g of the desired compound (36%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example δ.

Example 6δ

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-

2-methoxyiminoacetaraido]-3-[(l-amino-4-hydroxypiρe- ridinium)methyl]-ceph-3-era-4-carboxylate

By following the procedure in Example 64 except that 0.42g(3.6waol) of l-amino-4-hydroxypiperidine was employed in place of 1-amino- piperidinϋ, there was obtained 0.65g of the desired compound (42%).

The NMR spectra of the desired compound were the same as those of

8δ

the .compound prepared in Example 4.

Exaraple 69

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-y])- 2-methoxyiminoacetaraido]-3-[(2-amino-l,2,3,4-tetrahydro- isoquino1in-2-ium)methy1]-ceph-3-em-4-carboxy1ate

By following the procedure in Example 64 except that 0.53g(3.6ramol) of 2-araino-l,2,3,4-tetrahydroisoquinoline was employed in place of 1-arainopiperidine, there was obtained 0.78g of the desired compound < ^8% >-

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 5.

Exaraple 70 Synthesis of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)- 2-methoxyiminoacetaraido]-3-[(l-amino-l-raethyl-4- piperaziniura)raethyl]-ceph-3-em-4-carbox late

By following the procedure in Example 64 except that 0.55g(3.6mmol) of l-araino-l-methyl-4-piperazinium was employed in place of 1- aminopiperidine, there was obtained 1.14g of the desired compound (75%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 11.

Exaraple 71

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(4-amino-l,2,4-triazol-2-ium) methy1]-ceph-3-em-4-carboxy1ate

A) In 5ml of N.N-dimethylforraaraide were dissolved 2.52g(3.0mmol) of diphenylmethyl (6R,7R)-7-[(Z)-2-(2-tritylaminothiazol-4-yl)-2-methoxy- irainoacetamido]-3-chloromethy]-ceph-3-era-4-carboxylate prepared in Preparation Example 5 or 6 and 1.35g(9.0mraol) of sodium iodide and 0.34g (4.0mraol) of 4-amino-l,2,4-triazole was added thereto. The reaction mixture was stirred at 45—50'C for 3 hours and cooled to room temperature. 15ml of water was added, the reaction mixture was extratced with dlchloromethane and the organic layer was washed with water. The dichlororaethane layer was dried over anhydrous magnesium sulfate, concentrated under the reduced pressure and subjected to silica gel chromatography (5% methanol/dichlororaethane). The fractions containing the desired product were concentrated under the reduced pressure to give 1.71g of diphenylmethyl (6R,7R)-7-[(Z)-2-(2- tritylaminothiazol-4-yI)-2-methoxyiminoacetamido]-3-[(4-amin o-l,2,4- triazol-2-ium)methyl]-ceph-3-em-4-carboxylate iodide(56%).

B) To 1.6ral of anisole was added 0.56g(0.55mmol) of the compound prepared in the above A), 5.5ral of trifluoroacetic acid was slowly added under ice-cooling and the mixture was stirred for 1 hour. Diisopropyl ether was added, and the resulting precipitate was filtered and dried. A small amount of water was added, the pH was adjusted to 4.5—5 with saturated sodiura bicarbonate solution and the solution was subjected to HP-20 column chromatography using δ0% aqueous acetonitrile as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 0.21g of the desired compound ⁽ 78%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 12.

Example 72

Synthesis of <6R,7R)-7-[(Z)-2-(2-aminothiazoi-4-yl)-2- methoxyiminoacetamido]-3-[(1-amino-l,2,4-triazol-4-iura) methyl]-ceph-3-ea-4-carboxy1ate

By following the procedure in Exaraple 71 except that 0.34g(4mraol) of 1-araino-l,2,4-triazole was employed in place of 4-amino-l,2,4-triazole, there was obtained 0.63g of the desired compound(44%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 13.

Exaraple 73

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminotetrazol-4-iura) methy1]-ceph-3-em-4-carboxy1ate By following the procedure in Example 71 except that 0.34g(4mraol) of 1-arainotetrazole was eraployed in place of 4-araino-l, ,4-triazole, there was obtained 0.6g of the desired compound(41%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 16.

Exaraple 74

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(2-amino-l, 2,3, -tetrahydroisoquinol in-

2-ium) methy l]-ceρh-3-em-4-carboxy late In 6ml of dry dlchloromethane was suspended 0.59g(1.3mmol) of (6R,

7R) -7- [(Z) -2- (2-aminothiazol -4-yl)-2-methoxy irainoacetamido ] -3- acetoxyraethyl-ceph-3-em-4-carboxylic acid, 1.21ml (6. δmraol) of N,0-

bistrimethylsilyl trifluoroacetamide was added thereto under gaseous nitrogen atmosphere and the mixture was stirred at room temperature for 1 hour. 0.5ml(3.5ramol) of iodotrimethylsilane was added, and the resulting mixture was stirred at room temperature for 30 min. and concentrated under the reduced pressure. The residue was dissolved in 6ral of dry acetonitrile, 0.9ml of dry tetrahydrofuran was added thereto and the mixture was stirred for 10 min. 0.27g(2mmol) of 2-araino-l,2,3,4-tetrahydroisoquinoline and 1ml of dry dichlororaethane were added and the reaction mixture was stirred at room temperature for 3 hours. After completion of reaction, 0.3ml of water was added under cooling. The resulting precipitate was filtered, washed with a mixed solvent of acetonitrile and diethyl ether, dried and suspended in a small amount of water, after which the pH was adjusted to 5—-6 with saturated sodium bicarbonate solution. Then, purification was effected by HP-20 (Mitsubishi) column chromatography using δ% aqueous ethanol as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and -freeze-dried to give 26δmg of the desired conpound(3δ%) .

The NMR spectra of the desired compound were the sane as those of the compound prepared in Example 5.

Exaraple 75

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- aethoxyiainoacetaraido]-3-(1-aminoindoliniuamethy1)- ceph-3-ea-4-carboxylate

By following the procedure in Example 74 except that 0.27g(2araol) of

1-aminoindoline was eraployed in place of 2-amino-l,2,3,4-tetrahydro-

isoquinoline, there was obtained 0.23g of the desired compound(34%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 6.

Exaraple 76

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminobenzotriazol-3-ium) methyl]-ceph-3-em-4-carboxylate

In 6ml of dry dichlororaethane was suspended 0.59g(1.3ramol) of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetamido]- 3- acetoxymethyl-ceph-3-em-4-carboxylic acid, 1.21ral(6.5mmol) of N,0- bistrimethylsilyl trifluoroacetamide was added thereto under gaseous nitrogen atmosphere and the mixture was stirred at room temperature for 1 hour. 0.5nl(3.5mmol) of iodotriraethylsilane was added, and the reaction mixture was stirred at room temperature for 30 min and concentrated under the reduced pressure. The residue was dissolved in 6ml of dry acetonitrile, 0.9ml of dry tetrahydrofuran was added thereto and the mixture was stirred for 10 min. 0.296g(2.2mmol) of 1-araino- benzotriazole and 3ral of dry dichlororaethane were added and the reaction mixture was stirred at room temperature for 5 hours. After completion of reaction, 0.3ml of water was added under cooling. The resulting precipitate was filtered, washed with a mixed solvent of acetonitrile and diethyl ether, dried, suspended in a small amount of water and the pH was adjusted to 4—4.5 with saturated sodiura bicarbonate solution. -Purification was effected by HP-20 (Mitsubishi) column chromatography using 15% aqueous acetonitrile as an eluent. The fractions containing the desired product were concentrated under the

reduced pressure and freeze-dried to give 255mg of the desired compound (37%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 18.

Exaraple 77

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(l-aminobenziraidazol-3-iura) methyl]-ceph-3-em-4-carboxylate By following the procedure in Example 76 except that 0.29g(2.2ramol) of 1-aminobenzimidazole was eraployed in place of 1-aainobenzotriazole, there was obtained 0.29g of the desired compound(42%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 19.

Exaraple 78

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(1-amino-l,4,5,6-tetrahydropyrira idin- 3-ium)methyl]-ceph-3-em-4-carboxylate By following the procedure in Example 76 except that 0.22g(2.2mmol) of 1-amino-l,4,5,6-tetrahydropyrimidine was eraployed in place of 1-arainobenzotriazole, there was obtained 0.2g of the desired compound (31%).

The NMR spectra of the desired compound were the same as those of the compound prepared in Example 21.

Preparation Example 7

Synthesis of 4-raethoxybenzy] 7/5-(2-hydroxybenzylidenearaino)

-3-ch1ororaeth 1-ceph-3-em-4-carboxy1ate

In 50ral of methanol was dissolved 4.05g(10mraol) of 4-raethoxybenzy1 73-amino-3-chloromethyl-ceph-3-era-4-carboxylate hydrochloride, and 1.28ml(12ramol) of salicylaldehyde and 1.39ml(lOramol) of triethylamine were added thereto. The reaction mixture was stirred for 5 hours. The resulting precipitate was filtered, washed with 50ml of methanol and dried to give 4.26g of the desired compound(90%) . NMR (DMSO-dό, δ )

3.7 (d, 2H, C-2), 3.77 (s, 3H, 0CH ₃ )

4.53 (s, 2H), 5.22 (s, 2H)

5.35 (d, IH), 5.66 (d, IH) , 6.85—7.58 (m, 8H, phenyl

8.75 (s, IH)

Preparation Exaraple 8

Synthesis of 4-raethoxybenzy] 7 ?-(2-hydroxybenzylideneamino) -3-[(l-aminobenzotriazol-3-iura)methyl]-ceph-3-era-4-carboxy late iodide To 50ral of acetone was added 4.73g(10ramol) of 4-methoxybenz l 7/3- (2-hydroxybenzy1ideneamino)-3-chIororaethy1-ceph-3-em-4-carb oxy1ate prepared in Preparation Example 7 and 1.6g(10.7ramol) of sodium iodide was added thereto. The mixture was stirred for 3 hours in the darkness, concentrated under the reduced pressure to remove acetone. The concentrate was dissolved in acetonitrile(4.δral) , and 1.07ml (lOramol) of salicylaldehyde and 4.3g(30mraol) of 1-arainobenzotriazole were added thereto and the mixture was stirred at 20—25 ^* 0 for 24 hours. To the

reaction solution was added 50al of acetone and the insoluble was filtered off. A mixture of diisopropyl ether (500al) and diethyl ether (500al) was added to the filtrate. The resulting precipitate was filtered and dried to give 6.28g of the desired compound (90%).

3.76 (s, 5H, 0CH ₃ , C-2), 5.27 (s, 2H)

5.43 (d, IH, C-6), 5.δ2 (d, IH, C-7)

5.9 (q, 2H, C-3), 6.δ—7.6 (m, βH, phenyl)

8.0—8.33 (ra, 4H), 8.δ3 (s, IH)

Preparation Exaraple 9

Synthesis of 4-raethoxybenzy] 7 β -amino-3-[ (l-aminobenzotriazol-

3-ium)methyl]-ceph-3-em-4-carboxylate iodide

To 25ral of acetonitrile was added 7g(10mmol) of 4-methoxybenzyl 79-(2-hydroxybenzylideneamino)-3-[(l-arainobenzotriazol-3-iu m)methyl]- ceph-3-em-4-carboxylate iodide prepared in the above Preparation Example 8, and 10ml of cone.hydrochloric acid was slowly added thereto at 0—• 5°C. To the reaction mixture was added 100m] of isopropyl alcohol, and the resulting precipitate was filtered and dried to give 5g of the desired compound (80%). NMR (DMS0-d ₆ , )

3.74 (s, 5H, OCH3, C-2), 5.23 (bs, 211)

5.4 (s, 2H, C-6, C-7), 5. 95 (q, 2H, C-3)

6.8—7.3 (m, 411, pheny), 8.0—8.27 (m, 4H)

Preparation Example 10

Synthesis of 4-raethoxybenz 1 (6R,7R)-7-[(Z)-2-(2-aminothiazol-

4-yl)-2-methoxyiminoacetamido]-3-[(l-aminobenzotriazol-3- ium)methyl]-ceph-3-em-4-carboxylate iodide

To a mixture of phosphorus oxychloride (1.21ml, 12mmol) and ethyl acetate (40ml) was added 2.21g(llmmol) of (Z)-2-(2-aminothiazol-4-yl)- 2-methoxyiminoacetic acid and the mixture was stirred at 2—6°C for 30 min. 1.12ml(12mraol) of further phosphorus oxychloride was added and the mixture was stirred at 4—6°C for 30 min. 0.93ml (12mmol) of N,N-dimethylforraaraide was added and the mixture was stirred at the same temperature for 1 hour. Thus obtained solution was added to a solution of 6.31g(10ramol) of 4-methoxybenzyl 73-amino-3-[(l-aminobenzotriazol-3- ium)methyl]-ceph-3-em-4-carboxylate iodide prepared in Preparation Exaraple 9 and 14.8ml(60mmol) of N.O-bistrimethylsilyacetaraide in 40ral of ethyl acetate at -10—-5°C and the mixture was stirred at -5 ^e C for 30 min. To the reaction solution were added lOOral of ethyl acetate and 50ml of water and the pH was adjusted to 6—7 with sodium bicarbonate. The organic layer was dried over anhydrous magnesium sulfate, concentrated under the reduced pressure and 300ml of diisopropyl ether was added. The resulting precipitate was filtered and dried to give 5.δg of the desired compound (75%).

3.2δ—3.65 (m, 5H, C-2, 0CH ₃ ), 3.9 (s, 3H, OCH3) 5.1 (d, IH), 5.27 (bs, 2H) , 5.6—6.0 (m, 3H) 6.8—7.3 (m, 5H, phenyl, thiazole-H) 8.0—8.3 (a, 4H)

Exaraple 79

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-

- methoxyiminoacetaraidoJ-3-[(l-aminobenzotriazol-3-iura) methy]]-ceph-3-em-4-carboxylate

To a mixture of trifluoroacetic acid (15mi) and anisole (7.5ml) was added 7.8g (lOmraoJ) of 4-raethoxybenzy1 (6R, 7R)-7-[(Z)-2-(2-aminothiazol -4-yl)-2-methoxyi[«inoacetamido]-3-[(l-arainobenzotriazol-3 -iunι)Bietlιy]]- ceph-3-em-4-carboxylate iodide prepared in Preparation Exaraple 10 and the mixture was stirred at 35°C for 4 hours. The reaction solution was concentrated under the reduced pressure and added to 500ml of diethyl ether. The resulting precipitate was filtered, water was added thereto and the pH was adjusted to 4—4.5. Purification was effected by HP-20 column chromatography using 12% aqueous ethanol as an eluent. The fractions containing the desired product was concentrated under the reduced pressure and freeze-dried to give 3.9g of the desired compound (75%). The NMR spectra of the desired compound were the same as those of the compound prepared in Example lδ.

Preparation Exaraple 11

Synthesis of 7 -amino-3-[(l-aminobenzotriazol-3-iura)raethy1]-ceph- 3-em-4-carboxylate

In a mixture of trifluoroacetic acid (15ml) and anisole (7.5ml) was dissolved 6.3g(10mmol) of 4-methoxybenzyl 7/9 -araino-3-[(l-arainobenzo- triazol-3-ium)methyl ]-ceph-3-em-4-carboxylate iodide and the solution was stirred at 30—-35°C for 4 hours. The reaction solution was concentrated under the reduced pressure, 500ral of diisopropyl ether was added, and the resulting precipitate was filtered and dried. To the solid was added water and purification was effected by HP-20 column

9δ

chromatography using 15% aqueous ethanol as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 2.6g of the desired compound(75%). 3.7 (m, 2H, C-2), 5.2 (ra, 2H, C-6, C-7) 6.0 (q, 2H, C-3), 7.9—8.4 (m, 4H)

Exaraple 80

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetamido]-3-[(l-aminobenzotriazol-3-ium) methyl]-ceph-3-em-4-carboxylate To a mixture of phosphorus oxychloride (1,12ml, 12mmol) and ethyl acetate(40ml) was added 2.21g(llmraol) of (Z)-2-(2-arainothiazol-4-yl)-2- methoxyiminoacetic acid and the mixture was stirred at 2—6°C for 30 min. Further 1.21ml(12mraoI) of phosphorus oxychloride was added and the resulting mixture was stirred at 4—6"C for 30 min. To the reaction solution was added 0.93ml(12mmol) of N.N-dimethylforraamide and the mixture was stirred at the same temperature for 1 hour.

In 40ral of ethyl acetate were dissolved 3.5g(10ramol) of 7 _/ 3-araino-3- C(l-arainobenzotriazol-3-ium)methyl]-ceph-3-em-4-carboxy]ate prepared in Preparation Exaraple 11 and 14.8ml (60mmol) of N,0-bistrimethylsilyl- acetaraide, the solution prepared in the above was slowly added thereto at -10—5°C and the mixture was stirred at -5C for 30 min. After confirming the completion of reaction, 10ml of methanol was added and the solution was concentrated under the reduced pressure. To the residue was added water, the pH was adjusted to 4-—4.5 and purification was effected by HP-20 column chromatography using 12% aqueous ethanol as

an eluent. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried to give 3.7g of the desired compound(70%).

The NMR spectra of the desired compound were the same as those of the compound obtained in Example 18.

Preparation Example 12

Synthesis of 4-raethoxybenzy] (6R,7R)-7-[(Z)-2- (2-tritylaainothiazol-4-y])-2-methoxyiminoacetamido]-3- [(l-arainobenzotriazol-3-iua)methyl]-ceph-3-em-4- carboxylate iodide

In 15ml of dichlororaethane was suspended 0.63g(3mraol) of phosphorus pentachloride and the suspension was cooled to -30 ^* C. 1.2g(2.5mmol) of (Z)-2-(2-tritylaminothiazol-4-yl)-2-aethoxyiminoacetic acid hydrochlo- ride was added slowly and the mixture was stirred at -20—15°C for 2 hours. 1.63g(2.75mmol) of 4-methoxybenzyl 7 _/ 9-araino-S-Ed- arainobenzotriazol-S-iuii methy]]-ceph-3-em-4-carboxylate iodide obtained in the above Preparation Exaraple 9 was added to 30ml of acetonitrile, 2.22ml Omraol) of N,0-bistrimethylsilyl acetamide was added thereto and the mixture was stirred at 10—-15 ^* 0 for 1.5 hours. To this reaction solution was slowly added the solution obtained in the above at -30— -25°C, the raixure was stirred at -20—15 ^* C for 1.5 hours and 20ml of water and 40ml of ethyl acetate were added. The organic layer was washed with saturated sodium bicarbonate solution and with saturated brine. The organic layer was dried over anhydrous sodium sulfate and the solid was filtered off. The filtrate was concentrated under the reduced pressure, and dlchloromethane and diethyl ether were added.

The resulting precipitate was filtered and dried to give 1.68g of the desired compound(60%).

3.6—3.8 (a, 2H, C-2), 3.83 (s, 3H) 3.87 (s, 3H), 5.13 (d, IH), 5.3 (bs, 2H)

5.8—6.1 (a, 3H), 6.87 (s, IH, thiazole-H)

6.9—7.5 (a, 19H), 7.6—7.9 (ra, 4H)

Exaraple 81 Synthesis of (6R,7R)-7-[(Z)-2-(2-aainothiazol-4-yl)-2- methoxyiminoacetamido]-3-_.(l-aainobenzotriazol-3-ium) methyl]-ceph-3-em-4-carbo ylate

To l.Og(lmmol) of 4-methoxybenzyI (6R,7R)-7-[(Z)-2-(2-tritylamino- thiazol-4-yl)-2-methoxyiminoacetaraido]-3-[(l-aminobenzotria zol-3-ium) methyl]-ceph-3-em-4-carboxylate iodide prepared in the above Preparation Exaraple 12 were added 2ml of trifluoroacetic acid and 1ml of anisole, the mixture was stirred at 15—20°C for 2 hours and concentrated under the reduced pressure to remove trifluoroacetic acid. Diisopropyl ether was added, the resulting precipitate was filtered and dissolved in 5ml of methanol. The insoluble was filtered off and the filtrate was concentrated under the reduced pressure. A small amount of water was added to the residue, the pH was adjusted to 4—4.5 and purification was effected by HP-20 column chromatography using 12% aqueous ethanol as an eluent. The fractions containing the desired compound' were concentrated under the reduced pressure and freeze-dried to give 0.4g of the desired compound(75%).

The NMR spectra of the desired compound were the same as those of

the compound obtained in Exmaple 18.

Preparation Exaraple 13

Synthesis of diphenylmethyl (6R,7R)-7-[(Z)-2- (2-trityl minothiazol-4-yl)-2-methoxyiminoacctaraido]-3- [(4-amino-l, ,4-t iazo1-2-ium)methy1]-ceph-3-em-4- carboxylate iodide By following the procedure in Preparation Example 12 except that 1.62g(2.75mraol) of diphenylmethyl 79-araino-3-[(4-araino-l,2,4-triazol-2- ium)methyl]-ceph-3-em-4-carboxylate iodide was employed in place of 4-methoxybenzyl 73-amino-3-[(l-arainobenzotriazol-3-ium)methyl]-ceph-3- em-4-carboxylate iodide, tliere was obtained 1.78g of the desired compound (70%). 3.4—3.6 (m, 2H) , 3.78 (s, 3H) 5.2 (d, IH), 5.5—5.9 (ra, 3H) 6.8 (s, IH), 7.1 (s, IH) 7.1—7.6 (m, 25H), 8.5, 9.3 (s, s, 2H) Example- 82 Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- aethoxyiminoacetaraido]-3-[(4-amino-l,2,4-triazol-2-iura) methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 81 except that l.Og(lmmol) of the compound prepared in the above Preparation Exaraple 13 was employed, there was obtained 0.37g of the desired compound(78%).

The NMR spectra of the desired compound were the same as those of the compound obtained in Example 12.

Exaraple 83

Synthesis of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- yl)-2-carboxyraethoxyirainoacetamido]-3-[(1-arainobenzotriaz ol-

3-ium)methy]]-ceph-3-em-4-carbo ylate By following the procedure in Example 60 except that O.δgdramol) of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-carboxyme thoxyimino- acetaraido]-3-acetoxymethyl-ceph-3-era-4-carboxylic acid was eraployed in place of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-carboxyraethoxyirai no- acetaraido]-3-acetoxymethyl-ceph-3-era-4-carboxylic acid, there was obtained 0.15g of the desired compound(26%). MS(FAB, M+l) = 57δ NMR(DMS0-d ₆ + 20% DCl, )

3.8—4.0 (m, 2H), 4.73 (s, 2H) δ.l—δ.δ2 (m, 310, δ.82 (d, IH) 7.δ—δ.2 (m, 4H)

Example δ4

Synthesis of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- yl)-2-(2-carboxyprop-2-oxyiraino)acetamido]-3-[(l- aminobenzotriazol-3-ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Exaraple 49 except that 0.57g (1.09ramol) of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3-yl)-2-(2- carboxyprop-2-oxyimino)acetamido]-3-acetoxyraeth l-ceph-3-em-4-carboxy1ic acid was eraployed, there was obtained 0.12g of the desired compound (20%).

MS(FAB, M+l) = 603 NMR(DMS0-d ₆ + 20% DCl, δ )

1.3 (d, 610, 3.86—4.1 (m, 2JI) 5.05—5.55 (m, 3H), 5.8 (d, IH) 7.5—8.1 (a, 4H)

Example 85

Synthesis of (6R,7R)-7-[(Z)-2-(δ-amino-l,2,4-thiadiazol-3- y])-2-methoxyiminoacetaraido]-3-[(l-aminobenzotriazo]-3- ium)methyl]-ceph-3-em-4-carboxylate

In a mixture of acetonitrile (lral) and water (1.5ml) was dissolved 0.67g(5mmol) of lithium iodide and the solution was heated to 65—67 ^* 0. 0.24g(l.δmmol) of 1-arainobenzotriazole and 0.46g(lmraol) of (6R,7R)-7- [(Z)-2-(5-araino-l,2,4-thiadiazo]-3-yl)-2-methoxyininoacetam ido]-3- acetoxyraethyl-ceph-3-em-4-carboxylic acid were added and the mixture was stirred at 70°C for 1 hour while adjusting its pH to 6.8—7.1. After completion of reaction, the reaction solution was cooled to room temperature and stirred for 30 rain while adjusting the pH to 1—1.5 with 3N hydrochloric acid. The insoluble was filtered off, the filtrate was concentrated under the reduced pressure and subjected to column chromatography over aluminium oxide and over silica gel using δ0% aqueous acetonitrile as an eluent. The fractions were concentrated under the reduced pressure, dissolved in a small amount of water and purified by HP-20(Mitsubishi) column chromatography using 15% aqueous ethanol as an eluent. The fractions containing the desired product were concentrated under the reduced pressure and freeze-dried tσ give 0.14g of the desired compound in a pale yellowish white amorphous form (26.4%). MS(FAB, M+l) = 531

NMR(DMS0-d _ύ + 20% DCl , δ )

3. 6—4.0 (a, 2H) , 4.0δ (s, 3H) 5. 1 —5. 6 (m, 3H) , 5. δ5 (d, IH) 7.6— δ. l (ra, 4H)

Exaraple δ6

Synthesis of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- yl)-2-methoxyiminoacetaraido]-3-[(l-aminobenziraidazol-3- ium)methyl]-ceph-3-em-4-carboxylate By following the procedure in Example 85 except that 0.24g(l.δraaol) of 1-aminobenzimidazole was eraployed in place of 1-aminobenzotriazole, there was obtained 0.13g of the desired compound (24%). MS(FAB, M+l) = 530 NMR(DMS0-d ₆ , ) 3.3—3.7 (m, 2H) , 3. 8 (s, 3H)

5.0δ (d, IH), 5.4—5.85 (m, 3H) 7.5—8.δ (m, 4H), 10.1 (s, IH)

Exaraple 87 Synthesis of (6R,7R)-7-[(Z)-2-(5-aaino-l,2,4-thiadiazol-3- yl)-2-methoxyiminoacetamido]-3-[(1-amino-l,2,4-triazol-4- ium)methyl]-ceph-3-em-4-carboxylate

By following the procedure in Example 85 except that 0. lδg(l.δramol) of 1-amino-l,2,4-triazole was employed in place of 1-aminobenzotriazole, there was obtained 0.14g of the desired compound (29%). MS(FAB, M+l) = 481 NMR(DMS0-dd + 20% DCl, δ )

lOδ

3.25—3.7 (ra, 2H), 3.8 (s, 311) δ.l—5.3 (m, 3H), 5.6 (d, IH) 8.7, 9.7 (s,s, 2H)

Example 88

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminobenzotriazol-3-ium) methyl]-ceph-3-em-4-carboxylate sulfate

In 4ml of water was dissolved 0.5g(0.95ramol) of (6R,7R)-7-[(Z)-2- (2-aminothiazol-4-yl)-2-methoxyiainoacetamido]-3-[(l-aminobe nzotriazo]- 3-ium)methy]]-ceph-3-em-4-carboxylate prepared in Example 18, the solution was cooled to 5 ^* C and the pH was adjusted to 1—1.5 with 3N sulfuric acid. After stirring at the same temperature for 1 hour, 10ml of isopropyl alcohol was added and the mixture was stirred at 0—5°C for 4 hours. The resulting precipitate was filtered, washed with isopropyl alcohol and diethyl ether and dried to give 0.54g of the desired compound in a form of white crystalline solid (91%). m.p. = 176'C— (decomp.) MS(FAB, M+l) = 628

3.51 (q, 2H, C-2), 3.δ3 (s, 3H, OCHs)

5.2 (d, IH), 5.4—5.94 (m, 3H)

6.74 (s, IH, thiazole-H) δ.02—8.52 (m, 6H) , 9.6 (d, IH)

Example 89

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-y])-2-

methoxyiminoacetaraido]-3-[(4-amino-l,2,4-triazol-2-iura) methy1]-ceph-3-em-4-carboxy1ate su1fate

By following the procedure in Example δδ except that 0.2g(0.42ramol) of (6R,7R)-7-C(Z)-2-(2-arainothiazol-4-yl)-2-raethoxyirainoacet amido]-3- [(4-amino-l,2,4-triazol-2-ium)methyl]-ceph-3-em-4-carboxylat e prepared in Example 12 was employed in place of (6R,7R)-7[(Z)-2-(2-aminothiazol- 4-yl)-2-methoxyirainoacetamido]-3-[(l-aminobenzσtriazol-3-i um)raethy1]- ceph-3-em-4-carboxylate, there was obtained 0.22g of the desired compound (91.3%).

3.5 (ra, 2H, C-2), 3.δ3 (s, 3H, 0CH ₃ )

5.1 (d, IH, C-6), 5.4— .δδ (m, 3H, C-7, C-3)

6.75 (s, IH, thiazole-H), 8.76, 9.81 (s, s, 2H, triazole)

Exaraple 90

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2- methoxyiminoacetaraido]-3-[(l-aminobenziraidazol-3-iura) methy1]-ceph-3-em-4-carboxy1ate su1fate

By following the procedure in Example 88 except that 0.25g(0.47mmol) of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl)-2-methoxyiminoacetami do]-3-

C(l-aminobenzimidazol-3-ium)methyl]-ceph-3-em-4-carboxyla te Prepared in Exaraple 19 was employed in place of (6R, 7R)-7-[(Z)-2-(2-aminothiazol-4- yl)-2-methoxyiminoacetaraido]-3-[(l-aminobenzotriazol-3-iura )raethyl]-ceph- 3-em-4-carboxylate, there was obtained 0.26g of the desired compound (88%) .

3.3—3.6 (m, 2H, C-2), 3.δl (s, 3H, OCH3)

5.08 (d, IH, C-6) , 5. δ— 5. 9 (ra, 3H, C-7, C-3) 6.76 (s, IH, thiazole-H) , 7.5— δ . 4 (m, 4H, phenyl ) 10. 1 (s, IH, imidazole)

Exaraple 91

Synthesis of (6R,7R)-7-[(Z)-2-(2-aminothiazol-4-yl )-2- methoxyiminoacetamido]-3-[(l-aminobenzotriazo]-3-ium) methyl]-ceph-3-em-4-carboxylate maleate

In 1.5ml of water was dissolved 0.15g(0.2δmmol) of (6R,7R)-7-[(Z)- 2-(2-aminothiazol-4-yl)-2-methoxyirainoacetamido]-3-[(1-amin obenzotriazol -3-ium)raethyl]-ceph-3-era-4-carboxylate prepared in Example lδ, the solution was cooled to 5—lO'C, 0.034g(0.29amol) of maleic acid was added thereto and the mixture was stirred at δ ^* C for 2 hours. Λ mixture of isopropyl alcohol (12ml) and diethyl ether(3ml) was added and the resulting mixture was stirred at 0—5"C for 5 hours. The resulting precipitate was filtered, washed with diethyl ether and dried to give o.lδg of the desired compound in a form of white crystalline solid (δ2%). m.p. = 136°C— (decorap.) NMR (DMSO-dd, δ)

3.55 (q, 2H, C-2), 3.δ5 (s, 3H, 0CH ₃ )

5.19 (d, IH), 5.4—5.96 (a, 3H)

6.06 (s, 2H), 6.75 (s, IH, thiazole-H), 8.03—8.48 (a, 6H)

Example 92

Synthesis of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadiazol-3- yl)-2-methoxyiminoacetamido]-3-[(l-aminobenzotriazol-3-ium) methy1]-ceph-3-em-4-carboxylate suIfate By following the procedure in Example 86 except that 0.5g(0.94ramol) of (6R,7R)-7-[(Z)-2-(5-amino-l,2,4-thiadizaol-3-yl)-2-methoxyim inoacet- amido]-3-[(l-aminobenzotriazol-3-iura)raethyl]-ceph-3-era-4- carboxylate prepared in Example δ5 was employed in place of (6R,7R)-7-[(Z)-2-(2- aminothiazol-4-yl)-2-raethoxyirainoacetamido]-3-[(l-arainobe nzotriazo1-3- ium)methyl]-ceph-3-em-4-carboxylate, there was obtained O.δlg of the desired compound (δ6%). NMR (DMSO-dd, δ)

3.7δ—4.0 (ra, 2H), 4.03 (s, 3H) δ.lδ (d, IH), 5.4δ—5.9 (ra, 3H) 7.7—δ.05 (m, 4H)