JPS5930716B2 - Antibiotic manufacturing method - Google Patents

Description

DETAILED DESCRIPTION OF THE INVENTION The present invention relates to novel 3-thiolated-J me[acylamidocephalosporanic acid derivatives known to be effective as antibiotics.

This antibiotic is used as a supplement for animal feed, as a treatment for mastitis in livestock, and as a treatment for diseases caused by Gram-positive and Gram-negative bacteria in animals, including humans, and poultry. More specifically, the present invention relates to 3-thiolated-J-methyl-acylamide cephalosporins, where the 7-substituent is O-aminomethylphenylacetamide, O-aminomethyl-P
-hydroxyphenylacetamide or O-aminomethylphenylthioacetamide, the thiol group at the 3-position being 3-hydroxypyritazin-6-ylthiomethyl) and non-toxic pharmaceutically acceptable salts thereof; This invention relates to a method for producing easily hydrolyzable esters. 7
- Phenylacetamidocephalosporanic acid (7-AC
A(7)N-phenylacetyl derivative (also referred to as cephalolam, PACA and phenasporin) is well known in the academic literature.

Method for preparing the present compound (including those with and without substituents on the benzene ring, and also corresponding compounds in which the 3-acetoxymethyl group is substituted with methyl, hydroxymethyl and/or pyridinium methyl) and/or The following literature exists regarding properties. Chauvette,. R. R. Etc. Chemis
tryOfCephlOspOrinAntibiOt
ics. PreparationOnOfaNewClass
sOFA AntibiOticsandtheRelat
iOnOfStructureTOActivity'
2J0urna10fttheAmericanChem
ica1S0city184, 3401-3402 (
1 Chauvette,. R. R. etc. Struct
ureActivityOfRelatiOnship
sAnlOng7-AcylanaicephalOs
pOranicacid''AutimicrObial
Agents and ChemOtherapyl962
, 687-694. C0cker1J. D etc.? ? Cep
halOspOranicAcid. Part. Dis
placementOfAcetOxy-GrOupb
yNucheOphiles-JournalOfth
eChemicalSOcietyl5Ol5-503
1 (1966) Culp,. H. W. etc., ゛Metab
OlismandAbsOrptiOnOfthe7-
(Phenyl-AcetOamidO-1-Cl4)
-CephlOspOranicAcid'',.Ant
ibactarial Agents and ChemOt
therapy-1963, 243-2460C0cke
rsJ. D. etc., ゞゞCephalOspOranic
acidPart. 7-Acyl(MidOceph-
2-Em4-CarbOnyli・Cacids-JO
urnalOftheChemicalSOciety
lll42-1151 (1966).

JagO,. M. , ``AntibacterialAc
tivityOfSOmeDerivativesOf
7AminOcephlOspOrAnicacidA
gainstSlaphylOcOeusAureus
andSynergismBetweenthesea
nDOtherAntibiOticssslBritJ
．． PharmacOll22, 22-23 (1964)
.

LOderlB. , etc., FtTheCephalOsp
OrinC962).

Nacleus (71 AminOcephalOspO
ranicAcid)AndsOmeOfitsDer
tivatives″BlOchemicalJourn
al., 79408-416 (1961).

Nishida,. M. , etc., ゛StudiesOnM
icrObialDegradatiOnOfCeph
alOspOrinCDerivativesI[″,
TheJOllrnalOfAntiblOtics,
21.375-378 (1968) Nishida,.
M. , etc.TStudiesOfMicrObialD
egradatiOnOfCephalOspOrin
CDerivativesI-TheJournalO
fAntiblOtics, 165-169 (1968
). Spencer J. L. etc., 8Chemistry
y0fCepha10sp0rinAntibi0ti
cs. Synthesis and StructureA
cTivityRelatiOnshipOfCeph
lOspOudineAnalOguesAutimi
crObialAgeNtsandChemOtlle
rapy-1966, 573-580. S1edman
、． R. I. etc., 17- AmenOdesacelOxy'CepllalOs
pcranicacidanditsDerivati
ves゛J. MedChem, 7(1), 117-11.
9 (1964). Sullvan. H. Rl et al., ゛M
etabOlismOfOralCephalOthi
nandRelatedCepllalOspOrin
sintheRat″BlOchemicalJour
nal, lO2, 976-982 (1967). Vym
Ola. F. , etc., 8TheC1assificati
0nandCharacteristics0fCep
ha10sp0rinAntibiatiCsI. Sy
stemmaticStu 0ftheQuantita
tiveSensitivity0fS0mepath
0? NicMicrOOOrganismstOCeph
alcridine2'JournalOfHygie
neEpidemiOlOgy'MicrObiOlO
gyandlmmunOlOgyllOll8O-18
9 (1966) and numerous other patents on 7-acyl derivatives of 7-aminocephalosporanic acid.

For example, 7-[4-(α-aminoalkyl)phenylacetamide]cephalosporanic acid (U.S. Pat. No. 3,382,224)
1), 7-[P-aminophenylthio)acetamide]
-cephalosboranic acid (US Pat. No. 3,422,100), 7
mono(halophenylthioacetamido)cephalosporanic acid (U.S. Pat. No. 3,335,136) and an almost infinite number of derivatives represented by the general formula [Netherlan
d69/02013 (FarmdOc39, 174)]
etc., 7-(P-aminophenyl acetamide)-
Cephalosporanic acid is described together with its N-trityl derivative in US Pat. No. 3,422,103 [see also Japan 2712/67 (FarmdOc 25, 406)]. U.S. Patent No. 3,219,662 is R-CH2-C
Compounds in which R is phenyl, nitrochlorophenyl (especially P-nitro)chlorophenyl, alkyl phenyl and alkoxyphenyl] and the corresponding phenoxy-substituted compounds and the 3-acetoxymethyl group
- All derivatives substituted with a pyridinium methyl group are claimed.

There is also a wider range of them which includes a series of compounds in which R is phenylthio and compounds in which R is benzyl [i.e. 10 British patents for the use of compounds as starting materials, at least in general terms.
12943 and 1153421 (FarmdOc23
984) (UK Patent 1001474 and US Patent 32
80118).

U.S. Pat. No. 3,341,531 includes 71
-carboxamidomethylphenylacetamide) cephalosporanic acid and its betaines (Betaine)
s) is mentioned. US Patent No. 3,431,259
(FarmdOc27715) lists various 7-(halo-, dihalo-nitro- and halonitrophenylacetamido)cephalosporanic acids as starting materials for reactions with certain nucleophiles. Regarding 7-phenylacetamido)cephalosporanic acid having various substituents on the benzene ring, Japan 2717/67 (Farmd
Oc254O6), Japan 26105/69 (Farmd
Oc4O86O), UK 1178471 (FarmdO
c277l5, Netherlands 67/00906) and Japan 25785/69 (FarmdOc4O874).

Regarding the substitution of the 3-acetoxy group of cephalosporin with various heterocyclic thiols, see (a), (b), and (
c) is described in the literature group.

(a) South African Patent 70/2290 [see also Netherlands 70/05519 (FarmdOe8Ol88R), where the side chain is e.g.
- 3,4-thiadiazole-5-thiol and 1-methyl-1,2,3,4-tetrazole-5-thiol. (b) In the case of U.S. Patent No. 3,516,997, the side chain at position 7 is R3-(alk)m-CO-NH- and R3-S
-(Alk)m-CO-NH- (where R3 is one type of aromatic heterocycle), and the heterocyclic thiol at the 3-position is, for example, 1- Methyl-tetrazole 5-thiol and 2-methyl-1,3
・Includes 4-thiadiazole-5-thiol, etc.

(c) In US Pat. No. 3,563,983, compounds represented by the structural formula are obtained by reacting various cephalosporins, including cephalosporin C in some cases, with an aromatic mercaptan, which is a nucleophile.

In US Pat. No. 3,278,531 Ar is phenyl or certain substituted phenyls or certain aromatic heterocycles.

Similar nucleophiles such as 2-mercaptopyrimidine have been
3261832: UK, 1101422; USA, 347
9350: United States, 3502665 (all authorized by Gla.
xa).

A similar description can also be found in the UK, 1109525 (Ciba).

Nucleophiles of this type are described in other Fujisawa patents, namely Pelky 714518 (FarmdOc.
353O7; Netherlands 68/06129 and South Africa 2965/68), Canada 818501 (Farmd
Oc38845), UK 1187323 (FarmdO
c3l936; Netherlands, 67/14888) and especially the United States, 3516997 (FarmdOc34328;
The compound is called cefazoline and has a tetraazolyl acetyl side chain on the 7-amino group and a 5-methyl-thiadiazolylthiomethyl group at the 3-position. ing.

Regarding this compound, academic papers [e.g. Antimi
crObialAgentandChemOthera
pyl969, AmericanSOcietyLOr
MicrObiOlOgylBethesdalMar
yland P236-243 and J. AntiblO
tics (Japan) 23 (3) 131-148 (197
0)] was also reported. More recently, the substitution of the 3-acetoxy group of cephalosporins with various heterocyclic thiols has been described in US 3563983 and Netherlands 70/05519.

In this case, the side chain is, for example, 7-α-aminophenyl acetamide, and a typical heterocyclic thiol is 2-methyl-1
-3,4-thiadiazole-5-thiol and 1-methyl-1,2,3,4-tetrazole-5-thiol. And in the latter case this is U.S. Pat.
21 (filed on April 18, 1969, established on February 8, 1972). 35 other U.S. patents relating to similar statements
63983: Belgium JMo V1189 (FarmdOcl
28l7T), Japan 72/05550 (FarmdOc
l292lT) and Japan 72/0551 (Farmd
Ocl2922T). Formula and US, 3668203.

The following formula (where X includes -S-C- and -S-C-)
Cephalosporin having the structure shown in the above patent and US Pat. No. 3,239,515; 323,951
6;3243435;3258461;3431259
and 3446803.

Academic papers related to these are available in J. Med. Cheml
8, 174-181 (1965) and J. Chem.
SOc (LOndOn) 1595-1605 (1965
), 5015-5031 (1965), 1959-19
63 (1967).

There are various reports regarding the antibiotic methanesulfonate.

That is, Pelkey (742728 (FamdOe4l4
66R) describes the mono-methane sulfonate of ampinrin.

US Pat. Nos. 3,268,508 and 3,295,246 describe the conversion of one or more of the four amino groups of kanamycin to metasulfonate derivatives.

Sodium bacitracin methanesulfonate is described in US Pat. No. 3,205,137, and neomycin is described in J. AntibiOtics (Japan)
n) 12, 114-115 (1959). The injectable antibiotic collestimetate (COllsti
(commercially available as COly-Mycin by Warner-ChileOtt)
It is the methanesulfonate sodium salt of colistin.

Merklndex has 48 Japanese patents such as KOyama etc.
98 (1957) is cited. For colistimetate, please refer to the US Dispensat Ory 26th edition (J
．． B. LipincOtt, Philadelphia
A review is given in P315-320 of 1Penna). U. S. P. XVI
The 950 example is specific to the FPOlymyxin-formaldehyde sodium bisulfite compound. The present invention is based on the following formula (where X is H or OH and m is O or 1).

When X is 0H, m is O). The substances exist primarily in the zwitterionic form and their pharmaceutically acceptable salts (including dimethane sulfonate sodium salt) and easily hydrolyzable esters. The pharmaceutically acceptable salts mentioned above include the following.

i.e. non-toxic metal salts such as sodium, potassium, calcium and aluminum salts of carboxylic acids;
Similarly, non-toxic ammonium salts and substituted ammonium salts of carboxylic acids (examples of substituted amines include trialkylamines including triethylamine, procaine, dibenzylamine, N-benzyl-β-phenethylamine, 1-ephenamine, N-N'-di Benzylethylenediamine, dehydroabiethylamine, 1-ephenamine, N-N!-
bisdehydroabiethylethylenediamine, N-(lower)-alkylpiperidines (e.g. N-ethylpiperidine) and other amines used to form salts with benzylpenicillin) and non-toxic acid addition salts (i.e. amine salt). Examples of acid addition salts include mineral acid addition salts such as hydrochlorides, hydrobromides, hydroiodides, sulfates, sulfamates and phosphates and maleates, acetates, citrates, oxalate, succinate,
Acid addition salts of organic acids exist, such as benzoates, tartrates, fumarates, malates, mandelates, ascorbates, and the like. A particularly preferred salt is the dimethane sulfonate sodium salt of the compound of formula 1. These salts are water soluble and useful in the preparation of injectable drugs. Furthermore, the present invention includes amino groups such as t-butoxycarbonyl, carbobenzoxy, formyl, o-nitocafenylsulfenyl, β●β
・β-trichloroethoxycarbonyl, 4-oxo-2
Compounds (which may be intermediates or metabolic precursors) in which the amino group is protected by a substituent such as -pentenyl-2,1-carboethoxy-1-propenyl-121 Included.

Particularly common protecting groups are ketones (especially acetone) and aldehydes (especially formaldehyde and acetaldehyde) (e.g., U.S. Pat. Nos. 3,198,804 and 3).
347851) and β-ketoesters and β
- diketones (e.g. U.S. Pat. No. 7,332,5479) and β-ketoamides [Japanese Patent No. 71/24714 (Farm
dOc4732lS] etc.

The present invention also provides compounds of the formula: where X is H or 0H and m is O or 1 (with the proviso that if X is 0H, m is O); Provided are methods for making acceptable salts thereof and easily hydrolyzable esters thereof.

The manufacturing method is as follows. That is, a compound represented by the following formula or a salt thereof or an easily hydrolyzable ester thereof is combined with an acid represented by the following formula (where B is an amino protecting group, and the definitions of m and X are the same as the previous definition). The following formula (where B.X and m
(the definition of is the same as the previous definition) or a salt thereof or an easily hydrolyzable ester thereof,
Thereafter, the amino protecting group is removed to produce the compound of formula 1 or a pharmaceutically acceptable salt or easily hydrolyzable ester thereof.

The amphoteric compounds of the present invention are represented by the formula 3-thiolated-
J Aminocephalosporanic acid, i.e. 7-amino-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cepheme-4-carboxylic acid or its salts or easily hydrolyzable esters (US Pat. No. 3,284,451 and UK No. 1,229,453, U.S. Patent No. 324
Silyl esters used for 6-aminopenicillic acid in 9622 and UK 1073530, and pivaloyloxymethyl ester, acetoxymethyl ester, methoxymethyl ester, acetonyl ester, phenacyl ester, p-nitrobenzyl ester, β and β
・Prepared by coupling a compound of the formula (or a substance with the same function) as an acylating agent of a primary amine group to be done.

After coupling, the amino protecting group B is removed to obtain the desired product. Amino protecting group B can be used in peptide synthesis or 2
- Protecting group used in the synthesis of ampinrin, cephaloglycine, cephalexin, etc. from phenylglycine. Particularly effective protecting groups are protons as shown in the formula below, or β-diketones or β-ketoesters (US 3316247, e.g. methyl acetoacetate) or β-ketoamides [Japan, 71/24741 (
FarmdOc4732lS)] etc.

In the case of β-ketoamides, the acid having a protected amino group is preferably converted to a mixed anhydride with ethyl chloroformate or the like prior to reaction with the compound or its salt. After the coupling reaction the protecting group is removed to give the product of the invention.

For example, t-butoxycarbonyl groups are removed by treatment with formic acid or trifluoroacetic acid, carbobenzoxy groups by catalytic hydrogenation, and 2-hydroxy-1-naphthocarbonyl groups by acid hydrolysis. , the trichloroethoxycarbonyl group is removed by treatment with zinc powder in glacial acetic acid, and the 1-carboethoxy-1-propenyl group is removed by formic acid. It will be appreciated that other amino protections may be used and are within the scope of this invention. Equivalent substances for the acylating agent of the formula are acid halides and acid anhydrides (including mixed acid anhydrides).

In particular, mixed acid anhydrides prepared from lower aliphatic monoesters of carboxylic acids or stronger acids such as alkyl and aryl sulfonic acids and sterically hindered acids such as diphenylacetic acid are preferred. Furthermore, acid azides or active esters or thioesters (e.g. with p-nitrophenyl, 2,4-dinitrophenol, thiophenol, thioacetic acid) can be used, and the free acid itself can also be used, first by converting it to N-N'-dimethylchloro Fluorumiminium chloride [UK 1008170 and N
avak and Weichet, . Experiment
Coupling with the compound may occur after reaction with tiaXX [6, 360 (1965)].

In addition to the methods described above, methods for directly coupling free acids include the use of enzymes or N-mercarbonyldiimidazole or N-N'-carbonylditriazole [South African Patent Specification 63/2680, carbodiimide reagent].
Especially N. N/-dicyclohexylcarbodiimide N-N
'-Diisopropylcarbodiimide, N-cyclohexyl-N'-(2-morpholinoethyl)carbodiimide:
Sheehan and Hass,. J. Amer. Ch
em. SOcl 77, 1067 (1955)], alkylamine reagents [R. Buijle and H.
G. ViehesAngew. Chem. lntern
atiOnalEdditlOn3, 582 (1964
)], isoxazolium (IsOxasOl
ium) Salt reagent [R. B. WOOdward. R. A.
OlOfsOn and H. Mayerl J. Amer.
Clem. SOe. 83, 1010 (1961)] and the use of keteneimine reagents [C. L. Sle
vens and M. E. MOnd. J. Boat R. Chem
．． SOc. , 80, 4065]. Other acid equivalents are the corresponding azolade (AzOl
In other words, it is an amide of an acid in which the nitrogen atom of the amide forms a five-membered condensed aromatic ring having two or more nitrogen atoms.

Examples are imidazoles, pyrazoles, triazoles, benzimidazoles, benzotriazoles and substituted derivatives thereof. An example of a method for synthesizing azolade is as follows. That is, N-N'-carbonyldiimidazole is reacted with an equimolar amount of carboxylic acid in tetrahydrofuran, chloroform, dimethylformamide, or other inert solvent at room temperature to obtain a carboxylic acid imidazolide in quantitative yield. Carbon dioxide and 1 mol of imidazole are liberated in the process. The by-product imidazole is separated by precipitation and the imidazolide is isolated, but this is not very essential. Methods for synthesizing cephalosporin through these reactions and methods for isolating the produced cephalosporin are well-known methods. The above description is a method of coupling a free acid with a protected amino group to a compound using an enzyme.

This method involves adding esters of carboxylic acids such as methyl esters to various microorganisms [e.g. Takahashi et al., J.
．． Amer. Chem. SOc94al)4035-4
037 (1972) and T. Nara et al., J. Ant
iblOtjcs (Japan) 24jff). 5, 321-323 (1971)). After completion of the coupling and deprotecting reactions, the compound of formula I is converted, if desired, into a non-toxic, pharmaceutically acceptable salt or easily hydrolyzable ester thereof.

The following formula (where m and X are defined as above)
The zwitterionic product represented by is converted to dimethane sulfonate sodium salt represented by the following formula.

The conversion of the zwitterionic cephalosporanic acid into dimethane sulfonate is carried out in a known manner. A preferred method consists of reacting the zwitterion with sodium formaldehyde persulfite (or its raw materials sodium bisulsaite and formalin) and water in the presence of a strong sodium base (e.g. sodium 2-ethylhexanoate). The above mixture is heated at a temperature above room temperature (
For example, at 40° C. or higher, preferably 4045° C.), the solution is heated for a short time until a solution of the desired product is formed. The product can be prepared, for example, in a water-soluble anhydrous solvent (preferably ethanol,
(Alcohol such as isopropanol) is added to precipitate it as a solid and collect it. The present invention also provides a method for producing a compound represented by the following formula, a salt thereof, or an easily hydrolyzable ester thereof.

The method involves reacting 7-aminocephalosporanic acid or a salt thereof with a thiol represented by the following formula or a salt thereof, and, if desired, the resulting 7-amino-3(6-hydroxypyridazin-13-ylthiomethyl) This method consists of converting -3-cephame-4-carboxylic acid into its salt or easily hydrolyzable ester by a well-known method.

Ester groups (e.g. 3 of 7-aminocephalosporanic acid)
-acetoxy group) by a thiol group is a well-known reaction and is carried out in aqueous solution in the presence of a weak base such as sodium bicarbonate.

The preferred temperature for the substitution reaction is in the range of about 50°C-100°C. The novel 7-amino-3-(6-hydroxypyridazin-3-ylthiomethyl)-3-cephem-4-carboxylic acids of the present invention are used in the preparation of cephalosporin compounds of formula 1 or potentially useful derivatives thereof.

As is well known, the compound of formula 1 is prepared by first acylating 7-aminocephalosporanic acid or a salt thereof with the intended 7-side chain acylating agent, and then converting the 3-acetoxy group to 3-hydroxy-6-mercaptopyridazine. It can also be synthesized by another method in which the desired product is obtained by substituting with .

This alternative method is of course within the scope of the present invention. In the treatment of human bacterial diseases, the compounds of the present invention are administered into the bloodstream by conventional antibiotic administration methods.

The dosage rate is about 5-200 m'/K9 per day, preferably administered in divided doses of about 5-20 m'/K9 per day. They are administered in dosage units containing, for example, 125, 250 or 500 m9 of active ingredient (together with a physiologically acceptable carrier or excipient). Administration may also be in liquid form, such as a solution or suspension. The method for producing the starting material is shown below.

2-cyano-4-methoxyphenylacetic acid 12.77 (0.06 mol) of 4-methoxy-2nitrophenylacetic acid (D.G. Harvey et al., J. Che
m. SOc. , 1938, 97) and 2.47 (0
．． A solution consisting of 0.06 mol) of sodium hydroxide and 100 ml of water was dissolved at 50 psi hydrogen pressure at 10% of 17.
Hydrogenation was carried out over a palladium/activated carbon catalyst.

The theoretical amount of hydrogen was absorbed in 1 hour.

The catalyst was filtered and removed using a filter coated with diatomaceous earth (Celite). 4.27 (0.06 mol) of sodium nitrite (30 ml aqueous solution) was added dropwise to the slurry at 0° C., and then 20 ml of concentrated hydrochloric acid (0.25 mol) was added dropwise at the same temperature with stirring. Stirring was continued for ten minutes.
4.1y (0.03 mol) of potassium carbonate (10ml aqueous solution) was added to the reaction mixture.

Further, a mixed aqueous solution (33 ml aqueous solution) of potassium cyanide 107 and copper cyanide 7y was added at 0° C. with vigorous stirring. The mixture was stirred for ten minutes at 0°C, 1 hour at 10-15°C and finally 2 hours at 50°C. 3 of the product liquid
The solution was diluted with 00ml of water and filtered. The aqueous solution was acidified with concentrated hydrochloric acid and extracted with ethyl acetate (5 x 100 mO, then 5
% sodium carbonate solution (5 x 100 mO).
The aqueous extracts were combined, decolorized with a small amount of activated charcoal, cooled and acidified with concentrated hydrochloric acid, and extracted with ethyl acetate (3 x 100 ml). The extracts were combined with water (3 x 30 ml) and saturated brine (3
It was washed with ×30m0 and dried with anhydrous sodium sulfate.
Removal of solvent yielded 8.77 (76%) 2-cyano-4
-Methoxyphenylacetic acid was obtained, which was recrystallized from benzene-ligroin. M. P. l3O-132℃
H2-CO), 3.77 (3H,.s,.0CH3),
6.9-7.4 (3H..m phenyl H).

Analysis Calculated value (ClOH, NO3): C, 62.82
; Hl4.75; Nl7.33O actual value: Cl62.9
2; Hl4,74; Nl7.2O. 2-Aminomethyl-4-methoxyphenyl acetic acid hydrochloride8.
A solution consisting of 2y (0.043 mol) of 2-cyano-4-methoxyphenylacetic acid, 100 ml of ethanol and 50 ml of 6N hydrochloric acid was hydrogenated at 50 psi of hydrogen pressure over platinum oxide as a catalyst for 2 days.

The catalyst was separated. The sap was evaporated to dryness. The residue was washed with acetone (3 x 20 mO to give 5,57 2-aminomethyl-4-methoxyphenyl acetic acid hydrochloride. CH2-
CO), 3.70 (3H,.s,.0CH3), 3.9
(2H.q, 6.0H2, CH2-N), 6.5-7.
3 (3H,.m1 phenyl H), 8.5 (3H1br)
NII3+) 2-Aminomethyl-4-hydroxyphenylacetic acid 37 (0.013 mol)
48% of 4methoxyphenyl acetic acid hydrochloride and 40ml
The mixture consisting of hydrobromic acid was refluxed for 5 hours.

The reaction mixture was treated with activated carbon and neutralized (pH 6-7) with 6N sodium hydroxide to precipitate 2-aminomethyl-4-hydroxyphenylacetic acid. Collect a lot of this,
Washed with water and acetone and dried under reduced pressure. Yield 1.9
y (8 1.5%). M. P. >300'C
o2=t-butoxycarbonylaminomethyl-4-hydroxyphenylacetic acid2. 2 of IV (0.012 mol)
-aminomethyl-4=hydroxyphenylacetic acid and 1
．． 2.0 y of (
25 ml T'' solution of 0.014 mol) t-butoxycarbonyl azide was added and the mixture was diluted at room temperature with
It was stirred for 2 hours.

This was washed with ether, acidified with dilute hydrochloric acid and extracted with ether (2x150ml). The extracts were combined and diluted with water (2 x 150 ml) and saturated saline (3 x 50 ml).
Washed with m0 and dried. Removal of the solvent gave an oily residue, which was dissolved in chloroform-carbon tetrachloride (1:1).
, 5 Oml) and purification yields 1.7y (5 Oml).
2%) of 2-t-butoxycarbonylaminomethyl-4
-Hydroxyphenylacetic acid is a white solid (melting point 145-
147°C). Bu-H), 3.42 (2
H,. s,. CH2CO), 3.9 7(2H,d,
6 H2, CH2 N), 6.3 7- 7.1 (3
H,. m, phenyl H), 9.00 (IH) br NH
), 12. O(IH,.br NH).

Analysis, calculated value (C14H1905): C) 5 9.7
7; H) 6.81; N) 4.98o actual value: C)
58.84;H) 6.8 9:N) 4.7 9o Potassium o-[1-carbomethoxypropen-2-ylaminomethyl]-P-hydroxyphenylacetate 2
0.28 mol o-aminomethyl-p-hydroxyphenylacetic acid, 15.2 mol o-aminomethyl-p-hydroxyphenylacetic acid in a round bottom flask equipped with a 000 ml reflux condenser and an overhead stirrer.
8 y (0.28 mol) of KOH 6 4.9 6
V (0.56 mol) of methylacetoacetate and 1
50 Oml of pure methanol was charged.

The mixture was then refluxed for 3 hours. The solution was then concentrated to a small volume (100-150 mO, filtered and purified at 400-7 O
Diluted with 0 ml of anhydrous ether. The product crystallized by straining. Filter off the solid and vacuum desiccate overnight with P2O5
Dry on top. The yield of product (melting point 140-148°C) was about 80y. Sodium o -(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)-
P-hydroxyphenylacetate 0.6 y (0.
0 2 6 t -atoms) of metallic sodium and 5
An ethanolic solution of sodium ethoxide prepared from Oml of pure ethanol contains 0.026 mol of o-aminomethyl-P-hydroxyphenylacetic acid and 3.38 mol of o-aminomethyl-P-hydroxyphenylacetic acid.
One volume of ethyl acetoacetate (0.026 mol) was added and the mixture was refluxed for 6 hours.

The mixture was evaporated to dryness and the residue was recrystallized from ethanol to give about 6y of sodium o-(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)-p-hydroxyphenylacete as a colorless product. Obtained as needle-like crystals.
60 V (0.85 mol) of nitrous acid to a cooled solution of 100 y (0.76 mol) of t-butylcarbase, 87 y of glacial acetic acid and 120 ml of water. An aqueous sodium solution (5 Oml of water) was added dropwise over 40 minutes.

During this time, the temperature was maintained at 10-15°C. After the addition was completed, stirring was continued for an additional 30 minutes at the same temperature. 100 to this mixture
ml of water was added, and the separated oil layer was extracted with methylene chloride (
5×10 Oml). Combined organic extracts, 10
Oml of 10% aqueous sodium bicarbonate solution and 1o0m
The mixture was washed successively with 1 liter of water and dried over anhydrous sodium sulfate.
Methylene chloride was removed under reduced pressure over a water bath. The residual azide was distilled and collected at 45°C/2 OmmHg. 9 2
．． 71 (84%).

A mixture of 3-chloro-6-hydroxypyridazine (0.15 mol) of 3,6-dichloropyridazine and 5 Oml of acetic acid was refluxed for 2 hours.

The reaction mixture was cooled, diluted with 5 Oml of water and then evaporated to dryness under reduced pressure. Crystallize the residue from water
-Chloro-6-hydroxypyridazine with melting point 133-7
°C (literature value is 138-140 °C, N.Takabay
ashi,. YakugakuZasshi )7 5
, 778 (1955)) as a prismatic crystal.
5.8y (80%) was obtained. 3-hydroxy-6-
3- of mercaptopyridazine 2.67 (0.02 mol)
Chloro-6-hydroxypyridazine and 5.07(0
．． 07 mol) of freshly prepared potassium hydrogen sulfide and 30 ml of ethanol were heated in a sealed tube at 130-140° C. for 6 hours.

The reaction mixture was cooled and diluted with 200ml of water. Almost all the organic solvent was removed by vacuum distillation, and the remaining aqueous solution was acidified to pH 3 with dilute hydrochloric acid and extracted with ethyl acetate (
It was 6×50m0. The extracts were combined and evaporated to dryness. The residue was mixed with 307111 ethanol-ligroin (1:1
) to give 2.27 (87%) amorphous 3-hydroxy 6-mercaptopyridazine [melting point 158-15
9°C (literature value 157-158°C, J. Dnley et al., H
elv. Chem Actal 37, 121 (1954)
) was obtained. 3-Hydroxy-6-mercaptopyridazine A solution of 5.67 (0.05 mol) of 3,6-dihydroxypyridazine and 150 ml of pyridine was refluxed with 2.70y of pentasulfide phosphate under vigorous stirring. I added it little by little.

Refluxing was continued for 1 hour, then the reaction mixture was diluted with 200 ml of water and then concentrated to remove the pyridine. The resulting oily residue was suspended in water and extracted with ethyl acetate. The organic extracts were combined, concentrated again, and dispersed in a small amount of water to give 3-hydroxy-6-mercaptopyridazine as a yellow solid. Recrystallize from water to 0.62y (12
%) product (same appearance as above) was obtained. To a boiling solution of hydrazidihydrochloride of 3,6-dihydroxypyridazine 3157 (3 mol) and water of 21, 2957 (3 mol) maleic anhydride fine powder was added little by little with stirring.

Heating was continued for 4 hours after the addition was completed, and then left in the freezer overnight. 2857 (85%) of 3,6-dihydroxypyridazine were obtained.

M. P. 290 DEG C. 3,6-Dichloropyridazine A mixture of 1507 (1.33 moles) of 3,6-dihydroxypyridazine and 2507 phosphoric oxychloride was refluxed for 3 hours while avoiding contamination with water.

Excess phosphoric oxychloride was removed under reduced pressure and the dark residue was poured onto 1k9 crushed ice. A large amount of the generated precipitate was collected by filtration. The product was then also recovered by extracting the mother liquor 5 times with 300 ml of chloroform and treating it with 17 ml of activated carbon followed by evaporation of the solvent. The first and second recoveries were combined, dissolved in 500 ml of chloroform, then treated again with 17 activated carbon and further concentrated to yield 1657 (83%) of 3,6-dichloropyridazine with a melting point of 60-6FC ( ) was obtained as microneedles in a sealed tube. A suspension of 60y (0.4 mol) of 6-chloro-3-hydroxypyridazine in 3,6-dichloropyridazine and 200 ml of 10% hydrochloric acid was refluxed for 2 hours until a clear solution was obtained.

The clear solution was treated with approximately 1.5y of activated carbon and then filtered. The filtrate was concentrated under reduced pressure to give 6-chloro-3-hydroxypyridazine as colorless needles. Yield 4
9.57 (98%)M. P. l37l39°C 3-hydroxy-6-mercaptopyritadine507 (0.38 mol) of 6-chloro-3-hydroxy-pyridazine, 607
(0.38 mol) potassium hydrogen sulfide and 250 ml
A mixture of ethanol was heated in a 500 ml autoclave at 140°C for 14 hours with magnetic stirring.

The pressure reached 15-20k9/c. The mixture was evaporated to dryness and the residue was dissolved in 300 ml of water. The aqueous solution was acidified to pH 3 with dilute hydrochloric acid and added with 200 ml of ethyl acetate to pH 3.
Extracted 0 times. The combined extracts were concentrated to 34.37 (
70%) of amorphous 3-hydroxy-6-mercaptopyridazine was obtained. M. P. l5l-152℃potassium O-
[1-Carbomethoxypropene-2ylaminomethyl]
- Phenylthioacetate 0.28 mol O-aminomethylphenylthioacetic acid, 15,287 (0.28 mol) KOH and 64 ．．
967 (0.56 mol) of methyl acetoacetate and an additional 1500 m.p. E of pure methanol was charged.

The mixture was refluxed for 3 hours. Then add a small amount of the solution (1001
The mixture was concentrated to 50 ml), filtered and diluted with 400-700 ml of anhydrous ether. Crystals were precipitated by masatsu. The solid was filtered and dried over P2O5 in a vacuum desiccator. Sodium o-(1-ethoxycarbonyl-
1-propen-2-ylaminomethyl) phenylthioacetate 0.026 mol of o in an ethanol solution of sodium ethoxide prepared from 0.67 (0.0267 1 atom) of sodium metal and 50 ml of pure ethanol.
-aminomethylphenylthioacetic acid and 3.387(0
．． 026 mol) of ethyl acetoacetate was added and the mixture was refluxed for 6 hours.

The mixture was evaporated to dryness and the residue was recrystallized from ethanol to give sodium o-[1-ethoxycarbonyl-1propen-2-ylaminomethyl)phenylthioacetate as colorless needles. Methyl o-promomethylphenylacetate Methyl o-methylphenylacetate (
82.07, 0.50 mol), N-promosuccinimide (89.0V, .0.50 mol), benzoyl peroxide (
1.07) and carbon tetrachloride (800 mO) was refluxed for 2 hours under irradiation with a 750 Watt light source.

Succinimide was separated and the solvent was removed from the solution. The residue was vacuum distilled to yield 90.17 (74%) product. Boiling point 95-105°C (0.4mm); N. m. r(C
C4) Singlet 2.84 (4H), 5.50 (2H
), 6.31 (2H) and 6,38 (3H). Methyl 0-azidomethylphenylacetate o-promomethylphenylacetic acid (90.17, 0
．． 371 mol), sodium azide (26.0y,.0
．． 40 mol) and 10% acetone aqueous solution (750 ml
) was stirred at room temperature for 2 hours.

The solvent was removed under reduced pressure and the residue was dissolved in ether (300 mO
and water (100 ml). Crude methyl o-azidomethylphenylacetate (74.8) 7 obtained by drying and concentrating the ether solution was dissolved in 150 ml of methanol. This solution was ice-cooled and 150 ml of Treated with 3N methanolic sodium hydroxide. The mixture was left at room temperature for 1 hour, then evaporated to dryness, and the residue was then dissolved in water. The solution was acidified and the product was filtered off, dried and
Recrystallized from ethyl acetate-n hexane to give 49.57 (
70%) of the acid was obtained.

M. P. 116-118°C:N. m. r(CDCl3)
Sharp zinclet τ2.75 [4H], 5.63 (2H
) and nujiol 6.28 (2H); ν 21
00 and Maxl7OOcm-1.

Elemental analysis, calculated value (C9H, N3O2): C, 56.5
3:Hl4.75:Nl2l. 98.

Actual value: C, 56.37: Hl 4.65; N, 2l. 7
4OO-Aminomethylphenylacetic acid 0-azidomethylphenylacetic acid (9.67, 0.050 mol), 10% palladium on activated carbon (2.

57), methanol (150 mO and 1N hydrochloric acid (50 mO)
m1) was mixed with 30 p.m. s. Hydrogenation was carried out at i for 3 hours.

The mixture was filtered, concentrated under reduced pressure to approximately 30ml and extracted with ether. 1-2y starting material was recovered from the extract. Adjust the pH of the aqueous solution to 5.0 with dilute ammonia aqueous solution. The temperature was adjusted to O and cooled on ice. The white solid was collected by filtration, washed sequentially with ice water, methanol and ether, and dried in vacuo over P2O5. Yield 5.4f (65%
)M. P. l79-18FC (decomposition): N. m. r(C
F3CO2H): τ2.54(Sl4H), 5.48(
Ql2H) and 6.00 (Sl2H) 0-t-ptoxycarbonylaminomethylphenylacetate triethylamine (14.4y, 0,143 mol) were cooled on ice.

-aminomethylphenylacetic acid (10.3y, 0.062
4 mol) in an aqueous suspension (100 m0 of water, then t-
A THF solution (75 ml THF) of butoxycarbonyl azide (11.47, 0.080 mol) was added. The reaction mixture was stirred at room temperature for 16 hours and then diluted with THF under reduced pressure.
was almost removed. Wash the aqueous solution with ether and
Add 5ml of ethyl acetate, cool on ice, and adjust the pH to 3 with diluted hydrochloric acid.
．． Adjusted to 5. The ethyl acetate solution was dried and concentrated, and the solid residue was recrystallized from an ethyl acetate-n-hexane (1:1) solution. 14.47 (87%) white needles M.
P. 114-116°C was obtained.

Elemental analysis Calculated value (Cl4Hl9NO4): Cl63.
39, Hl 7.22: Nl 5.28.

Actual value: Cl63.44; Hl7.2l; Nl5.42
O2.4-dinitrophenyl o-t-butoxycarbonylaminophenyl acetate N-N5-dicyclohexylcarbodiimide (1.07, 0.0050 mol)
-t-butoxycarbonylaminomethylphenylacetic acid (
1.337, 0.0050 mol) and 2,4-dinitrophenol (0.927, 0.0050 mol) and 12 ml of anhydrous tetrahydrofuran.

The reaction mixture was kept at room temperature for 1 hour, then the precipitated N-
A large amount of N'-dicyclohexylurea was removed. The solvent was removed from the sap to obtain the active ester as a viscous yellow oil. O-t-butoxycarbonylaminomethylphenylacetic acid can be synthesized in quantitative yield from t-butoxycarbonyl azide and an amino acid by using triethylamine as a base.

BOC-amino acids react with thionyl chloride in the presence of triethylamine (solvent methylene chloride) or pyridine (solvent benzene) to give BOC aminoacyl chlorides.

This material can be coupled directly with a compound of the formula below, where R is as defined above, in the presence of triethylamine in methylene chloride.

The protecting groups may be removed by subsequent treatment with cold trifluoroacetic acid. Exactly 2007 7-aminocephalosporanic acid (7-ACA) was suspended in 500 ml of acetone and 2407 p-toluenesulfonic acid (5
00ml acetone) were mixed at once.

The mixture was stirred at room temperature for 5 minutes and then passed through a bed of diatomaceous earth (SuperCeF) and the bed was washed with 150 mf of acetone. Insoluble matter is about 3
It was 07. Next, 80 ml of water was added to the solution ▲, and while stirring, the inside of the flask was rubbed with a glass rod to crystallize p-toluenesulfonate. The suspension was stirred in an ice salt bath for 30 minutes and filtered cold. This was washed twice with 200ml of cold acetone (0°C) and air dried. 250y of salt was obtained.

This 7-ACA (7) p-toluenesulfonate is 21
Stir in methanol and remove insoluble matter with ``SuperCe''.
It was separated by l゛. The filtrate was placed in a 51-unit Mitsuro flask, and 21-units of water was added. The pH was then adjusted to 4 by adding concentrated aqueous ammonia while cooling, and the suspension was stirred at 0° C. for 1 hour. Collect the product by filtration and add 100ml of O'c water.
and 3 times with 11 parts of acetone (room temperature). The yield of 7-ACA after air drying was 1457. 0-Aminomethylphenylacetic acid can also be obtained by Beckman rearrangement of 2-indanone oxime and hydrolysis of the resulting lactam as shown below.

J. 0rg. Chem. 9, 380-391 (1944
) and 28, 2797-2804 (1963) operation 1
1000y of 2-interson oxime to 31600m
1 of chloroform at 20-25°C.

2 solution is cooled to -30°C (during this cooling process, some of the oxime crystallizes.

3 Add 14827 phosphorus pentachloride little by little to the vigorously stirred suspension.

The reaction temperature is controlled within the range of -28°C to -32°C by adjusting the rate of addition of solid phosphorus pentachloride. (-3
Best results are obtained at 0°C.

It is also possible to carry out the reaction at elevated temperatures from -10 to -5°C or perhaps even higher. However, this increases the production of tar and by-products and complicates the lactam isolation process. 4. Stir the reaction mixture at -30°C for 10 minutes after the addition is complete and then warm to 25°C over 3/4 hour.

During this time, solids dissolve and then new solids precipitate out.
5 The reaction mixture is stirred for a further 3 hours at 25°C and then 31600 ml of water are added to it at O-5°C with thorough mixing [the reaction can be followed by TLC (thin layer chromatography).

In a system consisting of 8 parts benzene and 2 parts acetic acid, the lactam is Rf
-0.36 and the oxime has an Rf of -0.64.
Spot is O. Color can be developed with 0.05% potassium permanganate spray. The oxime spots will not disappear completely, but they should become noticeably thinner. ) Washing is then carried out at 20-25°C. Separate the six phases and wash the chloroform phase with 15,800 ml of water.

7 Combine the aqueous phases and extract with 15800 ml of chloroform.

8. Combine the chloroform phases, add 15800 ml of water and adjust the pH to approximately 7 by adding 10% sodium hydroxide solution dropwise with thorough mixing.

This operation requires approximately 680 ml of 10% sodium hydroxide solution and the solution must be added gradually [
This base washing is important for removing by-products that produce tar. It takes 1-2 hours for the dropwise addition] 9. Separate the solution and wash the chloroform with 15,800 ml of water.

The 10 aqueous phases are combined, washed with 9200 ml of chloroform, and the chloroform portions are combined.

11 The chloroform solution was mixed with 1000 f7 activated carbon (゛Da
rkoKB') for 15-30 minutes at ~25°C.

12 Filter the slurry through "Dicalite", wash the cake with chloroform, and concentrate the slurry under reduced pressure.

The o-aminomethylphenylacetate lactam is then obtained as a dry solid. The yield of crude lactam 13 is almost 100%.

And it is a yellow crystalline solid. [If tar-forming substances have not been removed by washing, the product will be dark in color. This is first the slurry p
By adjusting H to 7.0, it can be recrystallized from hot water or from a toluene-heptane mixed solvent. ]Step 1 2s′10007 obtained by oxime rearrangement
Add 8000ml of concentrated hydrochloric acid to the crude lactam.

The two mixtures are carefully heated to reflux for 3 hours.

[Excess HCI evaporates during heating of the reactants and a large amount of foam is generated. This foam fills the entire device. This foam can be suppressed with silicone defoamers. Once the initial foam formation has ceased, the reactants can be refluxed without difficulty. ]3 Cool the dark slurry to about 40-50℃,
Add activated charcoal (“DarkOKB”) and continue stirring.

Activated carbon treatment was carried out for 415-20 minutes, and the slurry was
Callte゛The cake was filled with 40
Wash with 00ml of hot water.

5. Extract the clear yellow liquid with 6000 ml of methylene chloride and divide the methylene chloride.

The CH2Cl2 phase is saved to check for recovery of unconverted lactam.

6 Concentrate the aqueous phase under reduced pressure to obtain solid o-aminomethylphenylacetic acid hydrochloride.

7. Add MIBK to the wet solid and perform vacuum distillation.

MIBK is added until all water in the solid is removed.
Continue vacuum distillation until all 8MIBK is removed.

[MIBK distillation not only removes water azeotropically but also removes HCl at the same time. 9 Pour the solid again into 3900ml
of water and add 650ml (7) MIBK.

Adjust the pH of the solution to 5.0 with 6N ammonium hydroxide while stirring at 1020-25°C.

The o-aminomethylphenylacetic acid zwitterion has a pH of about 3.
Crystallization begins from step 5. 11 Stir the zwitterion slurry and cool to 0-5°C for 1 hour.

12 Pour the slurry and then boil the cake for ~1000m.
1 of ice water, 2000ml 1f) MIBKl then 5000ml
Wash carefully with ml of ice-cold acetone.

Combine the wash and wash solutions and check the lactam concentration. 13 The cake is aspirated and then dried in an air circulation open at 45°C.

The yield is 670-730y, which is 60-6 based on oxime.
It is 5%. 14 The reaction and process are followed by TLC using a solvent system of 5-acetone, 1-5 benzene, 1-O acetic acid, 1-5 water.

Color development is based on KMnO4. Rf (lactam) = 0.88
, Rf (amino acid) -0.69. O-aminomethylphenylacetic acid can also be synthesized by Schmiat rearrangement of 2-interson followed by hydrolysis as shown below.

0rganicSynthesis41, 53 (196
1) 2'-Intason is 0gganicSynthes
It is synthesized from indene by the method described in IS.

When industrially pure indene is used, the yield of 2-indanone decreases to ~45% (0rganieSy
see note 2 of nthesis). The 2-indanone obtained should be used as soon as possible. Because 0r
This is because, according to ganicSynthesis Note 7, 2-indanone is unstable in air at room temperature. Hydrolysis of the δ-lactam is carried out smoothly using hydrochloric acid to give amino acids. Yield 79%. Barium hydroxide may also be used, in which case the yield is 32%. A suspension of freshly prepared 2-indanone 12.3y (0.093 mol), sodium azide 13.07 (0.2 mol) and 500 ml of chloroform was cooled. Then, 50 ml of sulfuric acid is gradually added dropwise to the solution while stirring so that the temperature of the suspended liquid does not exceed 40'C.

After the addition is complete and the evolution of heat has ceased, the mixture is allowed to stand at room temperature for 2 hours.
Stir for an hour. Then pour over 4007 crushed ice. The chloroform phase is separated and the aqueous phase is extracted three times with 300 ml of chloroform. The chloroform phase and chloroform extract are combined, dried over anhydrous sodium sulfate, and concentrated under reduced pressure. Crystals were precipitated and collected by filtration to 250
Recrystallization from n-hexane-benzene (1:1) solvent yields 12.07 (88%) of the desired product as colorless prismatic crystals. M. P. l52-153℃IR(K
Br): νMax(C7n-1)320013050,
1660, 15001745.

NMR (CDCl3): δ (Ppm) 3.50 (2H,
．． t11.5Hz), 4.40(2H.br-S,.D
Converted to triflets by addition of 2O (J=1.5Nz
)), 7.06 (4H,.s), 7.45 (1H..b
(disappeared by addition of r-SlD2O).

Elemental analysis Calculated value (C9H9NO) ZC, 73.45:
Hl6. l6; Nl9.52O Calculated value: Cl73.73
;Hl6. O8; Nl 9.23 O-aminomethylphenylacetic acid δ-lactam 1.327 (0.01 mol) in a freshly prepared 21-Intason chloroform suspension (add 10 ml of sulfuric acid to 50 ml of chloroform with stirring) Then, 1.307 (0.02 mol) of sodium azide is added with stirring at room temperature.

After stirring for 30 minutes, the reaction mixture was poured into 200 ml of ice water and extracted twice with 100 ml of chloroform.

A large amount of insoluble matter generated during the extraction operation is separated. Combine the chloroform extracts, dry with anhydrous sodium sulfate,
Treat with activated carbon and filter. The filtrate is evaporated and the residue is recrystallized from 100 ml of alcohol-n-hexane (1:1) mixed solvent. 0.487 (32%) of δ-lactam with melting point 150-152°C is obtained. A mixture of 7.377 (0.05 mol) of o-aminomethylphenylacetic acid δ-lactam and 50 ml of concentrated hydrochloric acid was refluxed for 3 hours and then treated with 1.07 g of activated carbon. ,
I passed a lot.

The P solution was evaporated to dryness under reduced pressure to obtain a crystalline residue of 8.57. Recrystallization from 300 ml of acetone/water mixture (acetone:water=10:1) yielded 8.07 (79%) of the desired hydrochloride of the amino acid.

Colorless plate-like crystals. M. P. l64-166℃IR (KBr
): νMax(ClrL-1) 1695, 16101
1585, 1230, 11800N. M. R(DMSO
-D6): δ (Ppm from TMS) 3,69 (2H.
s), 3.94 (2H.s), 4.

10-4.7 (3H, .br0ad), 7.0-7.5
(4H,.m), 7.6-8.7 (1H.br0ad)
.

Elemental analysis Calculated value (C9HllNO2HCl): Cl5
3.6O:Hl6. OO:Nl6.95;Clll7.
58O actual value: Cl53.74; Hl5.98; Nl6
．． 79;Cll7.94. O-aminomethylphenylacetic acid δ-lactam of 340m9 (2.3mm) and 730η (2.3mm
A mixture of barium hydroxide octahydrate (01) and 20 WLe of water was heated at 150° C. for 1 hour in a sealed tube.

The reaction mixture was cooled to room temperature and heated to 300η (2.5mm01
) was added to precipitate the barium carbonate, which was filtered and thoroughly washed with 50 ml of water. The spring water was combined and evaporated to dryness. The residue was recrystallized from 5 ml of 50% aqueous alcohol to give colorless plate-like crystals of 163Tn9 (30%). This substance was confirmed to be the same as the free amino acid obtained by neutralizing amino acid hydrochloride with ammonium hydroxide. M. P. l8O-182°C. R
(KBr): νMax(C7rL-1), 310022
0011660-1500, 1400, 1380, 77
0,755,7200N. M. R. (CF3COOH)
:τ (Ppm from TMS) 3.96 (2H..br-
s), 4,45 (2H..brs), 6.3-8.0 (
3H. br), 7.38 (4H.s) Elemental analysis: Calculated value (C9HllNO2): C, 65.4
4; Hl 6.7l, Nl 8.48: Actual value: C, 65.
24: Hl6.4l:N, 8.3lO0-aminomethylphenylacetic acid lactam reflux condenser, drying tube, overhead stirrer,
24.67 (0.186 mol) of -2- into a 2 L three-liter flask equipped with a thermometer and a 100 ml dropping funnel.
Intason was charged with 26f7 (0.4 moles) of sodium azide and 1 liter of chloroform.

The suspension was stirred and 100 ml of concentrated sulfuric acid was added dropwise to it at a rate that maintained the temperature at 33-37°C (Note 1). After the addition was complete, the reaction mixture was stirred for an additional 2 hours at room temperature and poured into 8007 crushed ice. The chloroform phase was separated and the aqueous phase was extracted three times with 300 mj of chloroform. The chloroform extracts were combined, dried over anhydrous sodium sulfate, and then evaporated to dryness. The solid was redissolved in 150 ml of hot water and 2.0 f activated carbon (DarkoKB) was added.
It was heat-treated. The solution was filtered and the activated carbon was washed three times with 25 ml of hot water. When the P liquid was cooled, a solid precipitated, and the P was collected by filtration. Wash with 10-20 ml of ice-cold water,
Dry on P205 in a vacuum desiccator.

The product was 18.3f7 (69%), M. P144-14
It was 5℃. Note 1. Triazohydrogen acid is generated during addition of sulfuric acid.

Triazohydrogen acid boils at 38-39°C, so the temperature must be maintained below 40°C. Furthermore, all experiments must be conducted in a hood with sufficient ventilation. Note 2. Residual triazohydrogen acid in the aqueous phase can be removed by gradually adding sodium nitrite and dissolving it on iodine paper and/or dichloride.
Can be degraded by continuing until iron solution test is positive.

0-Aminomethylphenylacetic acid hydrochloride 500 equipped with reflux condenser and magnetic stirrer
10.1y (0.075 mol) o-aminomethylphenylacetate lactam and 100ml round bottom flask.
1 of concentrated hydrochloric acid was charged, and the mixture was refluxed for 3 hours.

While the reactants are hot, add 2.0 f7 activated carbon (“DarkoK
The mixture was treated with B) for 5 minutes and filtered thoroughly. 50-60℃
The mixture was concentrated and dried under high vacuum over a P205. (Note 1) The solid was recrystallized from an acetone-water (15:1) mixed solvent (Note 2). P the hydrochloride in a vacuum desiccator.
205. Pure product (M.P.188-19
The yield of "C) was 11.4y (78%). Elemental analysis Calculated value (C6H12NO2Cl): C, 53.73
;H1 5.97;N1 6,96;C1, 17.66
o Actual value C, 53.56; H, 6.02; N1 6.8
9; Cl1 17.76 Note 1 Completely dry material is required for successful recrystallization.

Note 2: The ratio of water and acetone varies depending on the degree of dryness of the crude hydrochloride.

Synthesis of o-aminomethylphenylacetic acid from indene 8
370ml 0% formic acid and 70ml 30% hydrogen peroxide
587 (0.5 mol) of indene was added dropwise to the mixture consisting of 587 (0.5 mol) at 35-40° C. over 1 hour while stirring.

An additional 40 ml of formic acid was used to wash away the indene from the addition funnel into the flask. The reaction mixture was heated at room temperature for 15 minutes.
I stirred for hours. The reaction mixture was concentrated to remove the formic acid under reduced pressure below 40°C (20-30 mm).

The residue was dissolved in 100 ml of 10% sulfuric acid, refluxed for 20 minutes and then steam distilled. Distillation was carried out at a rate of 21/hr until 71 distillates were obtained. 400% distillate
Extracted 6 times with 1 ml of chloroform. Integrate extracts,
It was dried over anhydrous sodium sulfate and then filtered. Evaporate the solution to dryness and obtain 50.27 (76%) of 2 with a melting point of 5657.
Obtained as needles at °C. 0-Aminomethylphenylacetic acid delta-lactam (3) 467 of the freshly prepared 2-indanone (2) (0.35 mol) in a cooled suspension consisting of 277 (0.42 mol) of sodium azide and 75 ml of chloroform. Add 75TfL1 of sulfuric acid to the solution at a temperature of 30
The mixture was added dropwise while stirring at a rate that maintained the temperature at -40°C.

Stirring was continued for 3 hours even after the addition was completed and heat generation stopped. The reaction mixture was poured into 1k9 crushed ice. The chloroform phase was separated and the aqueous phase was extracted three times with 200 ml of chloroform. The chloroform phase was combined with the chloroform extract, dried over anhydrous sodium sulfate, and evaporated to dryness. 44.5y (87%) of 3 was obtained as pale yellow prismatic crystals (M.P.l 46-149°C).

0-Aminomethylphenylacetic acid hydrochloride (4) 43.47
(0.30 mol) o-aminomethylphenylacetic acid δ-
A mixture of lactam (3) and 140TIII of 6N hydrochloric acid was refluxed for 4 hours.

The reaction mixture was treated with 1y activated carbon and evaporated to dryness. The residual oil was dispersed in 500 ml of acetone, purified, and
4 of f (75%) was converted into colorless plate-like crystals (M.P.l63-1
64°C). Synthesis of o-aminomethylphenylacetic acid from 0-nitrotoluene 130 ml (approximately 1 mol) of ethyl oxalate and 118
m1 (about 1 mol) of o-nitrotoluene (5) was added to 23
Pure ethanol solution of sodium (1f7 atoms) of y (
The mixture was added dropwise to 300 ml of ethanol while cooling.

The mixture was stirred at room temperature for 1.5 hours and then refluxed for 1.5 hours. The mixture was cooled to 60° C. and approximately 500 mi of water was added. The mixture was steam distilled to remove unreacted nitrotoluene. The residue was treated with 57 charcoal and filtered. The clear slurry was adjusted to pH 8-9 with dilute sodium hydroxide. 6% hydrogen peroxide was added at 30-40°C until no dark reaction of alkaline pyruvic acid was observed when a portion of the solution was sampled and made alkaline with sodium hydroxide.

The reaction mixture was treated with a small amount of activated carbon and filtered. The filtrate was acidified with concentrated hydrochloric acid and extracted four times with 250 ml of ethyl acetate. The extracts were combined, washed with water (3 x 100 m) and saturated brine, and dried over anhydrous sodium sulfate. The solvent was removed and recrystallized from ethanol/water (200 m - 600 m), 81.37 (45%) of o-nitrophenyl acetic acid (6) (melting point 140-14FC) was obtained.νCO
ll7lOcrfL-1, νNO2l525, 1360
-1 cm 0-Cyanophenyl acetic acid (7) A solution consisting of 43.4y (0.24 mol) of 0-nitrophenyl acetic acid (6) and 9.67 (0.24 mol) of sodium hydroxide aqueous solution (200 ml of water) was heated at room temperature with hydrogen Hydrogenation was performed using 1y of 10% palladium/activated carbon catalyst at 50 psi pressure.

The theoretical amount of hydrogen was absorbed in 4 hours.

Sawabetsu was the catalyst. While cooling the slurry to 0°C, 16.67 aqueous solution of sodium nitrite (50ml of water) was added.The mixture was added dropwise to 80ml of concentrated hydrochloric acid at 0-3°C with stirring.After the dropwise addition was completed, the mixture was cooled. An aqueous solution of potassium carbonate (100 ml water) of .67 (0.12 mol) was added to the reaction mixture with stirring at O-5°C. 40y (0.62 mol) of potassium cyanide and 287 (0.31 The diazonium salt was added to a solution of copper potassium cyanide (mol) of cuprous cyanide and 130 ml of water at O-5° C. with vigorous stirring.

The mixture was incubated at 0°C for 30 minutes, at 15-20°C for 1 hour, at 50°C.
The mixture was stirred for 1 hour and finally left at room temperature overnight. The reaction mixture was diluted with 1 liter of water and filtered. The filter solution was acidified with concentrated hydrochloric acid and extracted three times with 300 ml of ethyl acetate. The extracts were combined and back-extracted four times with 200 ml of aqueous sodium carbonate solution. The alkaline extract is treated with a small amount of activated carbon,
Acidified with concentrated hydrochloric acid and extracted three times with 300 ml of ethyl acetate. The extracts were combined, washed with 200 ml of water, 200 ml of saturated brine, dried over anhydrous sodium sulfate, and then concentrated to yield 367 crude product (1). Recrystallization from benzene gave 29.67 (77%) of a yellow colored product (M.P. 121-122°C). νc3N22
20CTL-1.

νc: 01690aπ~ 0-aminomethylphenyl acetic acid hydrochloride (4) (1) 1
．． 67 (0.01 mol) o-cyanophenyl vinegar 7)
A mixture of 2.57 rll of ethanol and 15 ml of 6N hydrochloric acid was hydrogenated at room temperature using a 1y 10% palladium/carbon catalyst.

A theoretical amount of hydrogen was absorbed for 18 hours.

The catalyst was removed and the sap was concentrated to give 1.6y (80%) colorless plate crystals. M. P. 161-163℃ This substance is synthesized by Schwidt rearrangement of 2-interson,
It was then identical to the true material obtained by hydrolysis.
(1) 3.2y (0.02 moles of 7, 0.5y of palladium/activated carbon, 50ml of ethanol and 25ml of
A mixture of 6N hydrochloric acid was hydrogenated at room temperature under a hydrogen pressure of 50 Pi.

The theoretical amount of hydrogen was absorbed in 3 hours.

The mixture was filtered to remove the catalyst, the filtrate was concentrated and the residue was purified with acetone (100 ml) to give 3.07 (70% of 4) (M.P. 162-163°C). . (i11
) A mixture of 3.22y (0.02 mol) of 7, 50 ml of ethanol and 25 ml of 6N hydrochloric acid was added to O.O.
Hydrogenation was carried out using platinum oxide of 15y as a catalyst at a hydrogen pressure of 50 PSi and a temperature of room temperature.

The theoretical amount of hydrogen was absorbed in 4 hours.

The catalyst was removed and the sap was evaporated to give a viscous oil. This was dispersed and purified in 150 ml of acetone to give a voltage of 2.1 V (
53%) colorless plate-like crystals (M.P. 161-163°C)
I got it. (1■ 7,20 of 1.6f (0.01 mol)
0.1f of platinum oxide was added to a solution consisting of ml of ethanol and 20ml of 6N hydrochloric acid and hydrogenated at normal pressure.

The theoretical amount of hydrogen was absorbed in 18 hours.

The catalyst was filtered off and the slurry was evaporated to dryness. 50ml of residue
Dispersion and purification in acetone of 1.07 (50%) of 4 (M
．． P. 161-163°C) was obtained. An aqueous solution of o-aminomethylphenylacetic acid hydrochloride containing 76.07 (0.378 mol) of 0-aminomethylphenylacetic acid (450ml of water was dissolved in 6N NH4
The solution was treated until reaching 0H' (pH 5).

The solution was cooled to 5-7°C and left overnight. A large amount of crystals was collected. 56.37 (92%), M. P. 188-190
°C elemental analysis Calculated value (C9H11NO2): C, 64
．． 45; H, 6.66; N, 8.48o actual value, C, 6
4.98; H1 6.86; N1 8.53.

Potassium o-[1-carbomethoxypropen-2ylaminomethyl]-phenylacetic acid 45.927 (0.28 mol) of o-aminomethyl was added to a 2000 ml round bottom flask equipped with a reflux condenser and an overhead stirrer. phenylacetic acid, 15.287 (0.28 mol) KOH,
64.96y (0.56 mol) of methyl acetoacetate and 1500 ml of pure methanol were charged.

The mixture was then refluxed for 3 hours. Then add a small amount (1
Concentrate to 0.00150mO, filter, 400-700mO
The mixture was diluted with 1 ml of anhydrous ether.

The product was then crystallized. The solid was filtered off and dried over P205 in a vacuum desiccator. (The solid material was extremely hygroscopic; it needed to be drained as quickly as possible; air drying was not possible) Product yield (M.P. 140-148 ℃) is 84.17
(99%).

Elemental analysis Calculated value (C14H16NO4K): C1 5
4.20; H1 5.49: N14.52: KF2.9
0o actual value: C, 53.72; H1 5.44; N14
．． 56; KF12.92.

Sodium o-(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)phenylacetate 0.
4.27 y(
One volume of o-aminomethylphenylacetic acid (0.026 mol) and 3.38 y (0.026 mol) of ethyl acetoacetate were added, and the mixture was refluxed for 6 hours.

The mixture was evaporated to dryness and recrystallized from ethanol to obtain 6.3
6t (82%) of sodium o-(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)
A portion of phenylacete is converted into colorless needle-like crystals (melting point 230-2
32°C). 1470, 1395, 1275,
1180α-1 o Elemental analysis Calculated value (C, 5H18N
O4Na):C) 60.2 0;H) 6.0 6:
N) 4.6 8.

Actual measurement; C) 59.95; H) 5.86; N) 4.
67ot-butoxycarbonyl azide 10 OV (7)
A solution of t-butylcarbazet (0.76 mol), 87 V glacial acetic acid and 12 Oml water was heated to 10-
60V (0.85 mol) over 40 minutes while maintaining at 15℃
An aqueous solution of sodium nitrite (5 Oml of water) was added dropwise.

After the addition was completed, the mixture was stirred for an additional 30 minutes at the same temperature. Add 100 ml of water to the mixture, and add 100 ml of the separated oil phase.
Extracted 5 times with 1 liter of methylene chloride. The organic extracts were combined and washed with 100 ml of 10% sodium bicarbonate solution,
It was dried with anhydrous sodium sulfate. methylene chloride 40-
It was removed under reduced pressure on a water bath maintained at 45°C. The residual azide was distilled and collected at 45°C/20 mmHg. The weight was 92.7 y (84%). o-(t-
butoxycarbonylaminomethyl) phenyl acetic acid 7 0
y (0.35 mol) of o-aminomethylphenyl acetate hydrochloride, and t-butoxycarbonyl azide in THF in a solution consisting of 116 V (1.15 mol) of triethylamine (TEA) and 4 Oml of water. A solution (THF 30 Oml) was added dropwise with stirring at 0°C.

After the dropwise addition was completed, the temperature was raised to room temperature and stirred for 20 hours. Tetrahydrofuran was distilled off at below 40°C, the aqueous solution was washed with 20 Omi of ether, and 2 ooml
of ethyl acetate was added and acidified to pH 3 with dilute hydrochloric acid at 0°C. The organic phase was separated and extracted four times with 20 Oml of ethyl acetate. Combine ethyl acetate and bring the solution to 20 Om
1 of water, dried over anhydrous sodium sulfate and concentrated in vacuo. The concentrate was treated with 500 ml (r)n-hexane to give an o-(t) of 87.9V (95%).
-butoxycarbonylaminomethyl)phenylacetic acid was obtained as colorless needle-like crystals (MP114 -116°C). 2,4-dinitrophenyl o-t-butoxycarbonylaminomethylphenylacetate o-(t-
butoxycarbonylaminomethyl) phenyl acetic acid (2
2.7 3 y) 0.0 8 6 mol), 2,4-dinitophenol (15.8 2 y) 0.0 8 6
dicyclohexylcarbodiimide (17.7 2 y) in a solution consisting of
086 mol) were added all at once.

The reaction mixture was stirred at room temperature for 2 hours, and the precipitated dicyclohexyl urea was separated and washed with 100 ml of T''. The filtrate and washings were combined and concentrated under reduced pressure below 50° C. to give a yellow oil. This was dispersed and purified in 150 ml of hexane to obtain 2,4-dinitrophenyl. -t-
Butoxycarbonylaminomethylphenylacetic acid was obtained as yellow needle crystals (M.P. 76-77°C). The yield was 34.9 y (94%). Elemental analysis, calculated value (C2o H2, N3 08 ): C, 5
5.6 8;H) 4.91;N) 9.4 7o actual measurement C) 5 5.7 0:H) 5.0 5:N, 9
．． 9 3o Preferred Specific Description The following examples are given by way of illustration and not as limitations of the invention.

All temperatures are expressed in degrees Celsius. 7-Aminocephalosporanic acid is abbreviated as 7-ACA.

That is, -ACA- represents a part having the structure, and in this way, 7ACA can be written as H-ACA-0-C-CH3.

Example 1 (Preparation of starting materials) 7-amino-3-(6-hydroxypyridazin-3-ylthiomethyl)-3-cephem-4-carboxylic acid 3-hydroxy-6-mercaptopyritadine 0.60y (0.0
047 mol) and 7-aminocephalosporanic acid 0.78
0.1 of y (0.0094 mol) of sodium bicarbonate
The mixture in 25 ml of M phosphate buffer (PH 6.4) was heated at 60° C. for 5 hours.

The reaction mixture was filtered to remove traces of insoluble matter, and the slurry was adjusted to pH 5 with acetic acid and produced a colored precipitate. This was collected by filtration, washed successively with water and acetone, and dried under vacuum to yield 1.037 (71 %) obtained.
Melting point 290-300°C (decomposition) 19Hz), 3.37 (1H, .d114Hz), 3.
65 (1H114Hz), 3.72 (1H,.d119
Hz), 4,90 (1H,.d14Hz), 5.30 (
1H. d, 4Hz), 6.75 (1H, d, 10Hz)
,7.30(1H,.d110Hz)Cl2Hl2N4
Elemental analysis value Cl4l calculated as O4S2・%H2O
．． 25, Hl3.75, N, 16.04S118,36
Experimental value Cl4l. 45, Hl3.7O, Nll5.8
3, Sll8. O3 Example 2 A7-(2-t-butoxycarbonylaminomethyl-4
-hydroxyphenylacetamide) 3-(6-hydroxypyridazin-3-ylthiomethyl)-3-cephem-4-carboxylic acid 1.77 (6 1,357 (6.6 mmol) of synchhexylcarbodiimide was dissolved in a solution of 1.2f (6.6 mmol) of 2,4-dinitrophenol and
was added all at once and the mixture was stirred at room temperature for 1.5 hours.

The separated urea was removed by filtration, and the filtrate was mixed with 3<em>1 water.
A solution of 2.07 (5.9 mmol) of 7-amino-3-(6-hydroxypyridazin-13-ylthiomethyl)-3-cephem-4-carboxylic acid dissolved in 1.7 ml (approximately 12 mmol) of triethylamine was added. Added all together. The mixture was stirred at room temperature for 20 hours and ethyl acetate (
The precipitate was washed with 3×150 mO and acidified with 6N hydrochloric acid, which was filtered, washed with water (50 m1), and dried. The precipitate was dissolved in methanol 11 at 50°C, and a small amount of insoluble matter was removed by filtration. The filtrate was treated with activated carbon and evaporated to dryness. When this was pulverized with ether (50 ml), 920~(25%) of 7-(2-t-butoxycarbonylaminomethyl 4-hydroxyphenylacetamide)-3-(6-hydroxypyridazin-13-ylthiomethyl)- 3-cephem-4-carboxylic acid was obtained. It had a melting point of 190-200°C (decomposed).
．． KBr IR:ν 17701170011670, Max
l58O, 1250? -1 The elemental analysis value calculated as C26H29N508S2.H2O is Cl5O. 23, Hl5. O3,N,ll. 2
7 Experimental value Cl5O. 63, Hl4.69, N, lO. 9
4 7-(2-aminomethyl-4-hydroxyphenylacetamide)-3-(6-hydroxypyridazine-3
-ylthio methyl)-3-cephem-4-carboxylic acid 3-7acetic acid (2m0 and 7-(2-t-butoxycarbonylaminomethyl-4-hydroxyphenylacetamide)-3-(6-hydroxypyridazine) 3-ylthiomethyl)-3-cephem-4-carboxylic acid (850η, 1
．． 4 mmol) were mixed while cooling to O-5°C and stirred for 1 hour.

Diluting the mixture with 150 ml of ether yields the desired cephalosporin, namely 7-(2-aminomethyl-4-
The trifluoroacetate of hydroxyphenylacetamide)-3-(6-hydroxypyridazin-3-ylthiomethyl)-3-cephemu-4-carboxylic acid is obtained. This was filtered and suspended in 30 ml of water. The suspension was adjusted to pH 6 with dilute aqueous ammonia and then diluted with 11 acetonitrile with stirring. The solid is collected, washed with acetonitrile (3 x 100a), and dried under vacuum to obtain the desired 7-
(2-Aminomethyl-4-hydroxyphenylacetamide)-3-(6-hydroxypyridazin3-ylthiomethyl)-3-cephem-4-carboxylic acid is 546 my
(77%) was obtained. Its melting point was 210°C (decomposed). KBr IR:ν 1770, 1660, 15801max
The elemental analysis value calculated as 1390C7rL-1 C21H21N506S2・3H20 is C, 43.82, H1 4.73, N1 1
2.56 experimental value C, 43.85, H, 4,28, N1
13.12 Example 3 7-(0-aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxypyrimidin-6-ylthiomethyl-3-cephem-4carboxylic acid N-N'-
To a stirred suspension of 2.1 mmol of sodium O-(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)-1P-hydroxyphenylacetate in 5 ml of dry acetonitrile containing drops of dimethylbenzylamine. Ethyl chloroformate O. with stirring at -15°C. 257
(2.3 mmol) and continued stirring at -10--15°C for 20 minutes.

To this mixture was added 0.717 (2.1 mmol) of 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-13-cephem-4-carboxylic acid and O. 21t (2.1 mmol) of acetonitrile 3m
1 and 3 ml of water are added together and the mixture is stirred at 0° C. for 1 hour. The reaction mixture is treated with a small amount of activated carbon and filtered. 0.5 d of formic acid is added to the filtrate while shaking, and the mixture is filtered to remove a small amount of unreacted 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid. Add 10% acetonitrile to the filtrate.
0 ml is added and the mixture is left at room temperature for 1 hour. The precipitate was filtered, washed with 107nj water, and vacuum dried to give 7-(
Approximately 0.5 of 0-aminomethyl-p-hydroxyphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid is obtained. Melting point: 210°C (decomposed) KBr IR: ν 1770, 1660, 1580, mo1
390, 1350, 1140, 1000crfl-1C
Elemental analysis value calculated as 21H21N506S2 (%
) is C1 43.82, H 4.73, N1 12.
56 Experimental value C, 43.85, H, 4.28, N1 1
3.12 Example 4 7-0-Aminomethyl-0-hydroxyphenylacetamide-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid Drying tube, overhead stirrer, and thermometer Got it, 1. Using a three-necked round bottom flask, add dry tetrahydrofuran (
Add well-ground sodium O-[1-carbomethoxypropen-2-ylaminomethyl]-P-hydroxyphenylacetate to 350 ml of THF) and four drops of N-N-dimethylbenzylamine.

The suspension is cooled to -40° in an acetone-dry ice bath with vigorous stirring. A suspension of 13.67 (0.06 mol) of isobutyl hechloroformate is added at once while maintaining the temperature below -35°. After 2.5 minutes, the acetone-dry ice bath was removed and 0.036 mol of 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3cephem-4-carboxylic acid and 5.0 mol of N-methylmorpholine were added. A solution of 0.047 (0.050 mol) in 250 ml of water is added.

The reaction mixture is stirred until it reaches room temperature (1-2 hours).
Remove T■゛ under reduced pressure. The aqueous solution is 100ml of ethyl acetate.
and acidified to pH 1.7-2.0 with concentrated hydrochloric acid. Add the mixture to activated carbon (“Darco KB”) 3.57
and filter. Separate the layers and add 1 part of the water layer to ethyl acetate.
Extract 00m 12 times. The water layer was then filtered through filter paper to a pH of 4. Adjust to O. After stirring vigorously,
The solution is left overnight for 5 to 7 days for crystallization. The product is collected by filtration, washed with a little water and dried in a vacuum dessicator over P2O5. 71 (0-aminomethyl-
The yield of P-hydroxyphenylacetamido)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid is about 30%.

Melting point 210°C (decomposition) Example 5 7-(0-aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxy-pyridazine-6-
ylthiomethyl)-3-cephem 14-carboxylic acid (
A zwitterionic suspension of 0.361y) in 3 ml of methanol is cooled with ice and a few drops of concentrated hydrochloric acid are added until a clear solution is obtained.

7-(0-aminomethyl-P-hydroxyphenyl*(
Acetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic hydrochloride hydrochloride precipitates as a light brown solid upon addition of ether, which is filtered and dried under vacuum over P2O5. .

Example 6 7-(0-aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxy-pyridazine-6-
ylthiomethyl)-3-cephem-4-carboxylic acid (0
．． 1 in a stirred suspension of the zwitterionic form of 3617).
Aqueous N sodium hydroxide solution was prepared as a clear solution (PHlO
．． Add until 8) is obtained.

Immediate lyophilization of this solution yields an impure solid, 7-(0-aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-14-carboxylic acid. Get sodium. Example 7 Method: 2.0 moles of 0-aminomethyl-P-hydroxyphenylacetamide-3-(3-hydroxy-pyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid (approx. 10107) and formaldehyde sodium sulfite 540f (4.03
mol) and 3000 ml of water and 30% SEH (sodium 2
-ethylhexanate) acetone solution 27007rL1
(4.87 mol) in a suitable container and heat the mixture to 40-45° C. while stirring.

The mixture dissolves into a yellow solution in about 10 minutes. heating 15
After a few minutes, decolorizing activated carbon (Darco KB) 507 was added to the solution and stirred for an additional 15 minutes at 40-45°C.

After a heating reaction at 40-45° C. for a total of 30 minutes, the mixture was filtered through diatomaceous earth (8 dicalide). 5 lumps of activated carbon
Wash with 2000ml of 0% ethanol water. Collect the filtrate 2
The temperature was adjusted to 5°C and the solution was stirred at high speed at 25°C.
0% ethyl alcohol. Add to file 1121. 71 (0-
A pure white amorphous precipitate with two sodium methanesulfonates attached to sodium aminomethyl-P-hydroxyphenylacetamide 3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylate is formed. .

Stir the suspension for about 10 minutes and then filter to remove 100% of the clumps.
Wash with ethyl alcohol 51. Dry the mass in a drier with air circulation at 50-55°C for about 2 hours, then dry it for 4 hours.
~6W! Dry under vacuum for 24 hours. What is obtained is 7-(0-aminomethyl-P-hydroxyphenylacetamide-3-(3-hydroxy-pyridazine-6)
-ylthiomethyl)-1-3-cephem-1-4-carboxylate is an amorphous, white solid with 1200 to 1400 y of sodium methanesulfonate attached. The product typically contains a few percent water and perhaps a small amount of ethanol. The product is 7-[0-N-N-bis(sodium sulfomethyl)aminomethyl-P-hydroxyphenylacetamide]-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem
-Also named sodium carboxylate.

Melting point Decomposes at 270°C or higher IR spectrum (KBr) 1760, 1680-1620, 1580, 1400,
120011040c7n-1 Hatakem spectrum (D2O
) 4.3ppm (4H,.s.N-CH2-SO3Na)
4,89 (1H.d, 4Hz, 6-H) 5.40 (1H
．． ．． d14Hz, 7-H) 6.78 (1H.d19.5
Hz1pyridazine-H)7.32(1H,.d19.5
Example 8 Prepare the following mixture: Formaldehyde Sodium sulfite 2.197 (2 equivalents) 7 (0-Aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxy-pyridazine 6)
-ylthiomethyl)-3-cephem-4-carboxylic acid zwitterion 3.57 (100-200 mesh) water 25
m1 (amount can be changed) 30% SEH-isopropanol solution 14dA nearly homogeneous solution is obtained by stirring vigorously at 24°C for about 0.5 hours.

Rapidly raise the temperature of the mixture to 40-43°C. This is allowed to stand for about 2 hours and then immediately cooled to 20-23°C. Filter the solution to remove some insoluble material (do not stay in solution for more than 2 hours).

The solution at pH 7.3 is added over 5 minutes to 600 ml of very rapidly stirring pure ethanol (other alcohols such as anhydrous isopropanol may be used as alcohol). 7-(0-aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-
An amorphous precipitate consisting of two sodium methanesulfonate molecules attached to sodium carboxylate is formed.

Stir the mixture for 5 minutes. Filter and collect the precipitate, 60 m
Wash with ethanol (or isopropanol) in step 1,
Vacuum dry at 50°C for 24 hours. The yield is about 4.3y0. The product has a pH of about 7 and is soluble in water to at least 200 m/ml.

The soybean solution is stable at room temperature for at least 2 hours;
It is stable for even longer periods in dilute solutions. The product exhibits the same antimicrobial spectrum as the starting zwitterion and is fully physiologically active whether or not it is hydrolyzed back to the zwitterion. Melting point Decomposes at 270℃ or above IR spectrum (KBr) 176011680-162011580114001
1200, 1040cm-1? Electric spectrum (D2O) 4.30ppm (4H, .s, .N-CH2-SO3N
a) 4.89 (1H.d, 4Hz16-H) 5.40 (
1H. d, 4Hz, 7-H) 6.78 (1H, .d19.5Hz1 pyridazine-H)
7,32 (1H..d19.5Hz1 pyridazine-H)
Example 97-[0-N-N-bis(sodium sulfomethyl)aminomethyl-P-hydroxyphenylacetamide]-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cefemu-4-carboxylic acid sodium.

Production method 7-1 using hydroxymethane sulfonate (0-
aminomethyl-P-hydroxyphenylacetamide)
Zwitterion of -3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid2.
05 mmol and sodium nitrium hydroxymethanesulfonate monohydrate 1.567 (10 mmol) and 1 MSEH
A mixture of 6 ml (6 mmol) of ethyl acetate solution, 10 ml of isopropanol and 10 ml of water is stirred at room temperature for 3.5 hours.

The reaction solution is treated with activated charcoal and poured into 300 ml of pure ethanol with stirring, and the mixture is stirred for 30 minutes at room temperature, resulting in a crystalline product. This was filtered and washed three times with 50ml of pure ethanol for 20 hours.
It is easily soluble in water when dried over P2O5 under the condition of 1f7/ml. Sodium 7-[0-N-N-bis(sodium sulfomethyl)aminomethyl-P-hydroxyphenylacetamide]-3-(3-hydroxypyridazin-6ylthiomethyl)-3-cephem-4-carboxylate is added to approx. Obtain 1.5y.

Melting point Decomposes at 270°C or higher IR spectrum (KBr) 176011680-1620, 1580, 14001
1200, 1040C7TL-1? m spectrum (D2
O) 4.30ppm (4H,.s,.N-CH2-SO
3Na) 4.89 (1H.d, 4Hz, 6-H) 5.4
0 (1H.d, 4Hz, 7-H) 6.78 (1H..d19.5Hz1 pyridazine-H)
7.32 (1H.d, 9.5Hz1 pyridazine-H)7
-[0-N-N-bis(sodium sulfomethyl)aminomethyl-P-hydroxyphenylacetamide]-3
-(Sodium 3-hydroxy-pyridazin-6-ylthiomethyl-3-cephem-4-carboxylate.

Manufacturing method using formalin and sodium sulfite 1 ml (10 mmol) of 30% formalin and 1 ml of sodium sulfite
7 dissolved in 10 ml of water, 7-(0-aminomethyl-P-hydroxyphenylacetamide)-
2 mmol of zwitterion of 3-(3-hydroxypyridazin-6-ylthiomethyl)3-cephem-4-carboxylic acid, 6 ml of 1M SEH solution and 10 me of isopropanol.
Add.

The mixture was stirred at room temperature for 2.5 hours and 300 m
Pour into. The resulting 7-[0-N-N-bis(sodium sulfomethyl)aminomethyl-P-hydroxyphenylacetamide]-3-(3-hydroxypyridazine-6
Sodium -ylthiomethyl)-3-cephem 4-carboxylate is collected by filtration, washed with three 50 ml portions of ethanol, and dried under vacuum. The yield is approximately 1657. )) 2 mmol of zwitterion of 7-(0-aminomethyl-P-hydroxyphenylacetamide)-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid and 1 MSEH ethyl acetate solution Add 1 ml (10 mmol) of 30% formalin to a mixture of 6 ml, 10 ml of isopropanol, and 10 ml of water.

The mixture is stirred at room temperature for 2 hours, resulting in a clear solution with a small amount of oily precipitate. After adding 17 of sodium sulfite, the solution is stirred for an additional 2 hours.

During this time, the oily precipitate dissolves into the solution. The reaction mixture was poured into 300 ml of ethanol with vigorous stirring to give approximately 1.87 7-[0-N-N-bis(sodium sulfomethyl)aminomethyl-P-hydroxyphenylacetamide]3-(3-hydroxy-pyridazine). Sodium 16-ylthiomethyl)-3-cephem-14-carboxylate was obtained, which was collected by filtration and 50 ml of ethanol was added.
Wash each piece three times and vacuum dry. Melting point: Decomposes at 270°C or above IR spectrum (KBr) 1760, 1680-1620, 158011400,
1,200, 1,040 accurate-1 spectrum (D2O) 4.30ppm (4H.s.N-CH2-SO3Na)
4.89(1H,.d14HZ16-H)5,40(1
H. d, 4Hz, 7-H) 6.78 (1H..d, 9.5Hz, pyridazine-H)
7.32 (1H.d19.5Hz1Pyridazine-H) Example 11A7-(0-Tert-Butoxycarbonylaminomethylphenylthioacetamide)-3-(3-Hydroxypyridazin-6-ylthiomethyl)-3-cephem-4- 2,4-dinitrophenyl ester of carboxylic acid BOC-protected amino acid 2.19f (4.7 mmol) was added to 5
0mlf) 1.027 (3 mmol) of 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid and 0.85ml of triethylamine were dissolved in THF at 0°C.
ml and add the solution dissolved in 20ml of water.

The reaction mixture is stirred at room temperature for 17 hours, treated with activated carbon and filtered. The filtrate is dried in vacuo to remove THF, and the concentrated product is washed five times with 100 d of ether each time. The aqueous solution is layered with ethyl acetate and acidified to PH2 with dilute hydrochloric acid at 0° C. with vigorous stirring. Add 100ml of ethyl acetate to the water layer.
Extract 3 times, one at a time. The organic layer, combined with the ethyl acetate extractant, is dried over anhydrous sodium sulfate and treated with activated charcoal. Evaporation of the filtrate under reduced pressure yields an oily product. When this was stirred and pulverized with 100ml of ether, 7-(0-Teat-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid was obtained as a pale yellow substance. Give as a solid. 10
The precipitate collected by filtration after washing with 0 ml of ether is 0.
．． 717 (38%) of the desired 7-(0-Teat-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid.

B7-(0-aminomethylphenylthioacetamide)
-3-(3-Hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid 7-(0-Tea
t-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid (0,
507, 0.8 mmol) is added in one portion to 3 ml of trifluoroacetic acid with stirring at 0°C.

Stirring was continued for 1 hour at 0-5°C. Addition of 100 ml of ether resulted in a white precipitate, which was filtered, collected and suspended in 4 ml of water at room temperature. The suspension was brought to pH 5 with dilute aqueous ammonia while stirring. When the precipitate is collected by filtration,
After washing with water and acetonitrile in this order and drying under vacuum over phosphorus pentoxide, a brown powder is obtained. Cefem-4-carboxylic acid was obtained. Weight is 220Tn9 (52%).
Melting point 200-210. C (decomposition). IR:νKBrl7
7O, 1670, 1580, Maxl2lOCTlL-
1.

1% NaHCO3 UV: λ 250nm (ε20800)
, ゜maxゝ270nm (ε14900) The elemental analysis values calculated as C2lH2lN5O5S3・2%H2O are C, 44.67, H, 4.64, Nll2
．． 4O, Sll7. O4 experimental value C, 44.43, H,
3.85, N, 10.84, S, 17.89 12-7-(0-Tert-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3 -Cephem-4-carboxylic acid 2,4-dinitrophenyl (0-Te
rt-butoxycarbonylaminomethylphenylthio)
Acetate 1.857 (4 mmol) (U.S. 3, 6
57, 232), 1.027 (3 mmol) of 7-amino-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid and 0.85 of triethylamine
m1 (approximately 6 mmol) in tetrahydrofuran (THF)
A solution of 30 ml of acetonitrile, 30 ml of acetonitrile and 50 ml of water was added while cooling to 0°C.

The mixture was slowly brought to room temperature and stirred for 12 hours. The reaction mixture was treated with a small amount of activated carbon and filtered. The organic solvent was removed from the filtrate by vacuum evaporation at below 35°C. The remaining aqueous solution was thoroughly washed with ether (30 ml x 4), a soil layer was made with 50 mj of ethyl acetate, and the pH was adjusted to 3 with dilute hydrochloric acid while stirring at 0 to 5°C. The precipitate formed between the two layers was collected by filtration. Its weight is 0.67. This was the desired 7-(0-Tert-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid. A concomitant amount of product was also obtained from the filtrate, which contained two layers. The aqueous layer was diluted with 10% ethyl acetate.
Extracted three times with 0ml. The organic layer was combined with the organic extractant, dried over anhydrous sodium sulfate, and evaporated to dryness. Wash the remainder thoroughly with 100ml of ether,
Vacuum dried over phosphorus pentoxide. Its weight was 0.427. The total yield of 7-(0-Tert-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid was 1.027 (55%). Ta.

Melting point 150-160°C (decomposed). IR:νKBrl77
O, 1690, 15901max1535, 1175C
T1-1 uv: λ1% NaHCO325OnWL (ε21700
), Max27O(Sh) (ε15500) The elemental analysis values calculated as C26H29N5O7S3・%H2O are Cl49.67, Hl4.97, Nlll.
l3, Sll5.3O experimental value C, 49.95, H, 4
．． 60, NllO. 7O, Sll5,52B 7-(0-aminomethylphenylthioacetamide)-
3-(3-hydroxypyridazin-6-ylthiomethyl)
-3-Cefem-4-carboxylic acid BOC-protected cephalosporin J Yoroi0 tert-butoxycarbonylaminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cefem-4-carboxylic acid 0.987 (1.58 mmol) and 4 ml of trifluoroacetic acid
The mixture was stirred at 0° C. for 1 hour.

Addition of 100 ml of ether produced a white precipitate, which was collected by filtration and washed with 100 ml of ether. Suspend the precipitate in 10ml of water and pH it with dilute ammonia.
When adjusted to 5, the desired cephalosporin Jyoroi0-aminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-16-ylthiomethyl)-13-cepheme-4-carboxylic acid is produced, which is filtered. It was collected, washed with 50 ml of acetonitrile, and dried under vacuum. Yield is 0
．． 46y (56%). It had the same IR spectrum as the product of Example 11 above. Example 13 7-(0-aminomethylphenylthioacetamide)-
2.1 mmol of sodium 3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid 0-(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)phenylthioacetate was added to -dimethylbenzylamine-drops of ethyl chloroformate into a stirred suspension in 5 ml of dry acetonitrile.
25y (2.3 mmol) is added with stirring at -15°C.

Stirring is continued for 20 minutes at -15°C. To the mixture 0.717 (2
, 1 mmol) and triethylamine 0.21y (2.1
A solution of 1 mmol) in 3 ml of acetonitrile and 3 ml of water is added all at once and the mixture is stirred at O'c for 1 hour.
The reaction mixture is treated with a small amount of activated carbon and filtered. Add 0.5 ml of filtrate heformic acid while shaking and filter the mixture to remove a small amount of unreacted 7-amino-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid. 100 ml of acetonitrile are added to the filtrate and the mixture is left at room temperature for 1 hour. The precipitate was collected by filtration, washed with 10 ml of water, and dried under vacuum to give 71 (0-
aminomethylphenylthioacetamide)-3-(3-
0.4f of hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid is produced. Similarly, when pivaloyloxymethyl ester is used instead of 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid, pivaloyloxymethyl 7- (0-aminomethylphenylacetamide)-3-(3-hydroxypyridazine-6
ylthiomethyl)-3-cephem-4-carboxylate was obtained.

Melting point 110℃ or higher (decomposition) IR spectrum (KBr) 17801 1750cT
rL-1NMR (DMSO) 1.15 (9H,.s),
5.11 (11H, .d14.5Hz), 5.60 (1
H. ．． d1 4.5Hz).

Example 14 7-0-Aminomethylphenylthioacetamide-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid Three 1 liter bottles with drying tube, overhead stirrer and thermometer 350 ml of dried tetrahydrofuran (THF) into a round-bottomed flask
ml, add four drops of 5N-N-dimethylbenzylamine,
Well-pulverized sodium O[1-carbomethoxypropen-2-ylaminomethyl]phenylthioacetate O. O8
Add moles.

The suspension is stirred vigorously and cooled to -40°C in an acetone-dry ice bath. Suspension isobutyl hechloroformate 1
Add 3,67 (0.06 mol) at once while keeping the temperature below -35°C. After 2.5 minutes, remove the acetone-dry ice bath and remove the 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cepheme-4-
Carboxylic acid 0.036 mol and N-methylmorpholine 5,
A solution of 0.047 (0.050 mol) in 250 mi of water is added.

The reaction mixture is stirred until it reaches room temperature (1-2 hours). THF is removed under reduced pressure. ethyl acetate 100ml
Separate the aqueous layer and add concentrated HCl to pH 1.7-2. Set it to O. The mixture is treated with 3.5 y of activated carbon (Darco KB) and filtered. Separate the layers and add the water layer to 100ml of ethyl acetate.
Extract twice each time. The aqueous phase was then filtered through filter paper and the pH
Adjust to 4.0. After stirring, the solution is left to crystallize overnight at 5-7°. The product is collected by filtration, washed with a small amount of water, and dried in a P205 vacuum desiccator. 7-
0-aminomethylphenylthioacetamide-3-(3
-hydroxypyridazin-6-ylthiomethyl)-3
The yield of -cephem-4-carboxylic acid is about 30%.

Example 15 7-(0-aminomethylphenylthioacetamide)-
3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid (0.361y)
A suspension of the zwitterionic form in 3 ml of methanol is cooled with ice and treated with a few drops of concentrated hydrochloric acid until a clear liquid is obtained.

Upon addition of ether, a precipitate of 7-(0-aminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic hydrochloride was formed as a light brown solid. , which is collected by filtration and vacuum dried on P205. Example 167-(0-
aminomethylphenylthioacetamide)-3-(3-
A 1N sodium hydroxide solution was added to a stirred suspension of the zwitterionic form of hydroxypyridazin-6ylthiomethyl)-3-cephem-4-carboxylic acid (0.3611) as a clear solution (pH 10) at room temperature.

Add until 8) is obtained.

When this solution is immediately freeze-dried, an impure solid 7-
(0-aminomethylphenylthioacetamide)3-(
3-Hydroxypyridazin-6-ylthiomethyl)-
Sodium 3-cephem-4-carboxylate is produced.
Manufacturing method example 17 of dimethane sulfonate Method: 2.0 mol of 0-aminomethylphenylthioacetamide-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cepheme-4-carboxylic acid (
anhydrous (approximately 1020 f), formaldehyde sodium nitrite 540 y (4.03 mol) and water 3000 ml and 3
2700 ml (4.87 mol) of 0% SEH (sodium 2-ethylhexanate) acetone solution is placed in a suitable container and the mixture is heated to 40-45° C. while stirring.

The mixture dissolves into a yellow solution in about 10 minutes. heating 15
After a minute, add decolorizing activated carbon (Darco KB) 50f7 to the solution.
and stir for an additional 15 minutes at 40-45K. Total 3
After a heating reaction at 40-45° C. for 0 minutes, the mixture is filtered using diatomaceous earth (dicalide). Wash the activated carbon mass with 2000ml of 50% ethanol and water. The filtrate was collected and adjusted to 25°C, and the solution was stirred at high speed at 25°C with 100% ethyl alcohol 1121? , add to. 7-(0-Aminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazin-16-ylthiomethyl)-3cephem-4-Sodium carboxylate with two sodium methanesulfonates, pure white amorphous A precipitate forms.

The suspension was stirred for about 10 minutes and then filtered to remove any clumps.
Wash with 151% ethyl alcohol. The mass is dried in a drier with air circulation at 50-55° C. for about 2 hours and then under a vacuum of 4-6 mm for 24 hours.

What is obtained is 7-(0-aminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazine-
6-ylthiomethyl)-3-cephem-4-sodium carboxylate with two sodium methanesulfonate atoms, amorphous, white solid 1200 to 1400
It is f7.

The product generally contains several percent water and may contain a small amount of ethanol. This product is 7-[0-
N-N-bis(sodium sulfomethyl)aminomethylphenylthioacetamide]-3-(3
-hydroxypyridazin-6-ylthiomethyl)-3
-Also named sodium cefem-4 monocarboxylate.

Melting point decomposed at about 250°C IR spectrum (KBr) 1770, 1670-1640, 1580, 1210 (
V7l-1 Example 18 Prepare the following mixture: Formaldehyde Sodium sulfite 2.19V (2 equivalents) 7-(0-aminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazine-
3.5 V (100-20
0 mesh) water 2 5ml (amount can be changed)
30% SEH-isopropanol solution 14 ml24
After stirring vigorously for about 0.5 hours at 0.degree. C., a nearly homogeneous solution is obtained.

Rapidly raise the temperature of the mixture to 40-43°C. This 2
Continue for a minute and then immediately cool to 20-23°C. Filter the solution to remove some insoluble material (do not stay in solution for more than 2 hours).

The pH 7.3 solution is added to 600 ml of very rapidly stirring pure ethanol in 5 minutes (other alcohols such as anhydrous inopropanol may be used as alcohol). Amorphous precipitate with two sodium methanesulfonate attached to sodium 7-(0-aminomethylphenylthioacetamide)-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylate is generated.

Stir the mixture for 5 minutes. The precipitate was collected by filtration and 6 O
Wash with 1 ml of ethanol (or isopropanol) and vacuum dry at 50° C. for 24 hours. The yield is about 400 g. The product is soluble in water at a pH of about 7 to at least 200 g.

Such solutions are stable at room temperature for at least 2 hours;
It is stable for even longer periods in dilute solutions. The product exhibits the same antimicrobial spectrum as the starting zwitterion and is fully biologically active whether or not it is hydrolyzed back to the zwitterion. Melting point: decomposed at about 250°C IR spectrum (KBr) 17 70) 16 70-16 4 0, 1580,
1210crrL-1 Example 19 7-[O-N-N-bis(sodium sulfomethyl)aminomethylphenylthioacetamide]-
Sodium 3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylate.

Production method 7 using hydroxymethane sulfone -(
0-aminomethylphenylthioacetamide)-3
-(3-Hydroxypyridazine-6-ylthiomethyl)-3-cephem-4-carboxylic acid Zw
itter ion 2.0 5 mmol and sodium hydroxymethanesulfonate monohydrate 1
．． 56V (10 mmol) and 6 mi (6 mmol) of IM SEH in ethyl acetate and 10 mi of isopropanol.
and 10 g of water is stirred at room temperature for 3.5 hours.

The reaction solution is treated with ly activated carbon, poured into 300 ml of pure ethanol with stirring, and the mixture is stirred for 30 minutes at room temperature, resulting in a crystalline product. This was filtered and washed 3 times with 50 ml of pure ethanol for 20 hours.
7-[0-N-N-bis(sodium sulfomethyl)aminomethylphenylthioacetamide]-3-(3 -Hydroxypyridazin-6-ylthiomethyl)-3-cephem-
Sodium 4-monocarboxylate is produced. Melting point: approx. 250
IR spectrum decomposed at °C (KBr) 1770, 1670-1640) 1580, 1210
clrL-1 Example 20 7-[0-N-N-bis(sodium sulfomethyl)aminomethylphenylthioacetamide]-3-(
3-Hydroxypyridazin-6-ylthiomethyl)-
Sodium 3-cephem-4-carboxylate.

Manufacturing method using formalin and sodium sulfite (a) 3
Add 1 ml (10 mmol) of 0% formalin and 17 ml of sodium sulfite dissolved in 10 ml of water in order.
-aminomethylphenylthioacetamide)-3-(3
-Hydroxypyridazin-5-ylthiomethyl)-3
-2 mmol and 1 zwitterion of cefem 4-carboxylic acid
Add 6 ml of MSEH solution and 10 ml of isopropanol.

The mixture was stirred at room temperature for 2.5 hours and 300 m
Pour into 1. The generated sodium 7-[0N-N-bis-(sodium sulfomethyl)aminomethylphenylthioacetamide]-3(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylate was collected by filtration. , washed three times with 50 ml of ethanol each time, and dried under vacuum. (b) 2 mmol of zwitterion of 7-(0-aminomethylphenylthioacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid, 6 ml of 1 MSEH ethyl acetate solution, and isopropanol. Add 1 ml (10 mmol) of 30% formalin to a mixture of 10 ml and 10 ml of water.

The mixture is stirred at room temperature for 2 hours, resulting in a clear solution with a small amount of oily precipitate. After adding 17 of sodium sulfite, the solution is stirred for an additional 2 hours.

During this time, the oily precipitate dissolves into the solution. The reaction mixture was poured into 300ml of ethanol with vigorous stirring to give 71[0-N-N-bis(sodium sulfomethyl)].
Aminomethylphenylthioacetamide]-3-(3-
A solid of sodium hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylate is obtained, which is collected by filtration, washed three times with 50 ml portions of ethanol, and dried under vacuum. Melting point decomposed at about 250°C IR spectrum (KBr) 1770, 1670-164011580, 1210c
TrL-1 Example 21 A7-(0-Tert-butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)3-cepheme-4-carboxylic acid 0-t-butoxycarbonylaminomethylphenylacetamide Dissolve 1.9y (3.75 mmol) of 2,4-dinitrophenyl enyl acetate in 15 ml (71) THF and add to the stirring solution while keeping the temperature at 0°C. A cooled solution of 1.147 (3.3 mmol) of )-3-cephem-4-carboxylic acid and 0.627 (6.6 mmol) of triethylamine in 15 ml of acetonitrile and 5 ml of water was added at once.

The reaction mixture was allowed to warm slowly to room temperature and stirred overnight. Remove the organic solvent from the mixture under reduced pressure below 30°C,
The remaining aqueous solution was washed with ether (10ml x 3).
A layer of 20 ml of ethyl acetate was formed on top of the water layer, and the pH was adjusted to 2.5 with dilute hydrochloric acid while stirring, and an organic layer was separated. The aqueous layer was extracted with ethyl acetate (100ml x 3). The insoluble material generated during the extraction was collected by filtration and washed with a small amount of water to form a pale yellow powder.
t-Butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)3-4-carboxylic acid (0.30f
7) was generated. (A). Those associated with organic extractants were treated with activated carbon and filtered. The filtrate was dried over anhydrous sodium sulfate and the solvent was evaporated to give an oily substance, which was thoroughly washed with 50ml of ether to obtain 0.59f of 71 (01Tert-butoxycarbonylaminomethylphenate) as a pale yellow powder. Enylacetamido)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem 4-carboxylic acid was produced (B). (A) (B) Both products showed the same spectrum. Total yield is 0.89
7 (46%).

Melting point 160-17'C (decomposition). KBr IR: ν 1780, 1710, 1690, Max
l68Oll53Oc7n-1 uv:λ1%NaHCO325Onm (ε18400)
MaxNMR: DMSO δ Ppm l. 35 (9H1 s), 4.02 (2H.s), 4,02 (2H,.s), 3.
3-4.7 (5H, unseparated), 4.95 (1H, Dl4
Hz), 5.50*(1H,.d-Dl9, 4Hz),
6.64 (1H, .d110Hz), 7.04 (4H.
．． s), 7,18 (1H, .d110Hz), 8.90
** (1H..d, 9Hz), 12.60** (1H wide single absorption).

(*Double line (J-4Hz) when D2O is included)**D2O
). C26H29N5O7S2・%H2
The elemental analysis values calculated as O are Cl52.34, Hl5
．． O7, Nllll. 74, SllO. 75 Experimental value: Cl
52. O3, Hl4.98, Nlll. 37.SllO
,O9B 7-(0-aminomethylphenylacetamide)-3-
(3-hydroxypyridazine-161ylthiomethyl)-
3-cephem-4-carboxylic acid 7-(0-Tert-butoxycarbonylaminomethylphenylacetamide)
-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid (500, 0.
87 mmol) was added to 3 ml of trifluoroacetic acid with stirring at 0°C, and the mixture was stirred for an additional hour below 0°C.

Ether (100ml) was added to the reaction mixture and the resulting precipitate was collected by filtration. The precipitate was suspended in 10Tn1 of water, and the pH of this suspension was adjusted to 5 with stirring with dilute ammonia water.
It also followed me. The desired product 7-(0-aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4
-The carboxylic acid was collected by filtration, washed successively with 5 ml each of water and acetonitrile, and dried in vacuo over phosphorus pentoxide. Yield: 286 soybeans (67%). Melting point is 222-2
30°C (decomposition). KBr IR:ν 177011665, 16401max
158011395, 1005CTrL-1uv:λ1
%NaHCO325On77L, Max (ε18400
), 270nm (Sh) (ε14600) NMR: δD2O-K2CO33.7h (2H.s),
Ppm4. l5(2H,.s), 3.0-4.

3 (4H1 unseparated), 4.90 (1H.d14Hz),
5.45 (1H..d14Hz), 6.63 (1H1d
19Hz), 7.15 (4H,.s), 7.20 (1H
．． ．． d. ．． Hz).

The elemental analysis values calculated as C2lH2, N5O5S2・1%H2O are Cl49, O2, Hl4.7l, Nll3
．． 6l, Sll2.46 Experimental value: Cl48.66, Hl
4.52, Nll3. O4, Sll2.43 Column 22 A7-(0-Tert-butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cepheme-4-carboxylic acid 0-Tert-butoxy Carbonylaminomethylphenylacetic acid 1.0y (3.7 mmol) and 2.4-
To a solution prepared by dissolving 0.697 (3.7 mmol) of dinitrophenol in 8 ml of dry ethyl acetate, 0.78 f of N-dicyclohexylcarbodiimide was added all at once.
The mixture was stirred at room temperature for 1 hour.

The separated synchlohexyl urea was filtered off and the filtrate was evaporated under reduced pressure to yield the activated ester as a yellow oil. The active ester was dissolved in THF3Od and cooled to 0°C. To the solution, combine a mixture of J-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid (1.02y13 mmol), triethylamine 0.85 ml, THF3Om, and water 10 ml. added. The reaction mixture was stirred at room temperature for 16 hours and treated with activated carbon. The filtrate was diluted with 50 ml of water and washed twice with 100 ml of ether. 100 on top of the water layer
A layer was formed with 1 ml of ethyl acetate and acidified to PH2 with dilute hydrochloric acid at 5-10° C. with vigorous stirring. The aqueous layer was extracted twice with 100 ml of ethyl acetate. The organic layer was combined with ethyl acetate extractant, dried over anhydrous sodium sulfate, treated with activated carbon, and filtered. When the solvent was evaporated, 7-(0-Tert-butoxycarbonylaminomethylphenylacetamide)-3- was obtained as a pale yellow oily solid.
(3-hydroxypyridazin-6-ylthiomethyl)-
3-cephem-4-carboxylic acid was produced. This was stirred and ground with 100 ml of ether. The BOC protected product was collected by filtration, washed with 30 ml of ether and dried in vacuo. Yield: 0.95y (54%). B7-(0-aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-
Carboxylic acid BOC protected cephalosporin i.e. 71 (
0-Tert-butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3cephem-4-carboxylic acid0
, 957 (1.6 mmol) and 6 ml of trifluoroacetic acid was stirred at 0-5° C. for 1 hour.

Ether (100ml) was added to the reaction mixture and the resulting white precipitate was collected by filtration and washed with 100ml of ether. The precipitate was suspended in 10 ml of water, and the pH was adjusted to 5 with dilute ammonia water. Desired product, 7-(0-aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4
- The carboxylic acid was filtered off, washed with 50 ml of acetonitrile and dried under vacuum over phosphorus pentoxide. Yield 0.50y (6
1%). Example 23A7-(0-t-butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxypyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid 0-t-butoxycarbonylaminomethylphenylacetamide 2・4-dinitrophenyl 93y was poured into 600 ml of tetrahydrofuran and cooled (-3 to 0°C
), add J-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4 to the stirred liquid.
-A cooled solution of carboxylic acid 627 (0.18 mol) in 250 ml of water was added in one portion.

The reaction mixture was stirred overnight and the temperature was allowed to reach room temperature. The mixture was treated with 107 activated carbon and filtered. The filtrate was concentrated to about 300 ml under reduced pressure below 40°C. Wash the concentrated solution with 5 times of 200ml of ether, and then wash it again with about 20ml of ether.
7 activated carbon. Add the ether of step 11 onto the filtrate and acidify with dilute hydrochloric acid to pH 3.5 while stirring vigorously to obtain crude 7-(0-t-butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxy Pyridazin-16-ylthiomethyl)-3-cephem-4-carboxylic acid is produced, which is collected by filtration and 21
The solution was washed with 21 parts of ether, 21 parts of water, and 0.51 parts of ether. Its weight was 20y. The crude product was divided into two parts and one portion (507) was used in the next step without further purification.

The other portion (40 y) was dissolved in 11 ml of methanol to remove a dark colored solid material (3.9 y), and the filtrate was concentrated to about 50 ml.

When 300ml of acetonitrile is added to the concentrated solution, the result is 17.4
A pale yellow precipitate of y was formed, which was collected by filtration and washed with 100 ml of acetonitrile. Concentration of the mother liquor gave an additional 12.57 units of product. S7-(0-Aminomethylphenylacetamide)-3-(3-hydroxypyridazin-16ylthiomethyl)-3-cephem-4-carboxylic acid (a) Crude 7-(0-Tert-butoxycarbonylaminomethylphenyl 507 mL of 3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid was dissolved in 100 ml of trifluoroacetic acid with cooling.

The solution was stirred at 0-5°C for 1.5 hours. When 11 ethers are added to the solution, trifluoroacetate of the desired cephalosporin is produced, which is collected by filtration and ether 300%
Washed with m1.

Dissolve trifluoroacetate in 100ml of water to make a 10% solution.
When the pH is adjusted to 4 with aqueous ammonia, a colored product (
26.

67) was produced, which was washed with 300 ml of water and 300 ml of acetonitrile in this order, and dried under vacuum.

The crude product was dissolved in 50 ml of formamide at room temperature, and when 150 ml of water was added dropwise to the solution, a small amount of colored precipitate formed, which was removed by filtration. When 1.5 l of acetonitrile was added to the filtrate, a pale yellow powder of 7-(0-aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4 was obtained.
-Carboxylic acid was produced.

Its weight was 9.870 (b) 7-(0-tert-butoxycarbonylaminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem, reprecipitated by the same method as described above. -4
- two parts of carboxylic acid (17,4y and 12.57)
to give the desired product 7-(0-aminomethylphenylacetamido)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cefemu-4-carboxylic acid 12.3y.

The two products from (a) and (b) above showed almost identical 1R spectra and MIC values.

This was analyzed together. Melting point: 208°C (decomposed). C2
The elemental analysis values calculated as 1H21N505S2/2%H20 are C147.36, H14.92, N1 13.
15, S1 12.04 Experimental value: C147.30, 47
．． 56, H14.37, 4.50,. N,. 13,67,13.
71, S1 12.38 Example 24 7-(0-aminomethylphenylacetamide)-3-
(3-hydroxypyridazin-6-ylthiomethyl)-
Sodium 3-cephem-4-carboxylic acid 0-(1-ethoxycarbonyl-1-propen-2-ylaminomethyl)phenylacetate O. 637 (2.1 mmol) as N-
0.25 f (2.3 mmol) of ethyl hechloroformate suspended under stirring in 5 ml of dry acetonitrile containing N'-dimethylbenzylamine droplets was added with stirring at -15°C, and the stirring was continued at -10 to -15°C. C. for 20 minutes.

Add 0.717 (2.1 mmol) of 7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid and triethylamine O to the mixture.
．． 217 (2.1 mmol) in acetonitrile 3TLe
and a solution dissolved in 3 ml of water were added all at once, and the mixture was heated to 0°C.
The mixture was stirred for 1 hour. The reaction mixture was treated with a small amount of activated carbon and filtered. Add 0.5 ml of filtrate heformic acid while shaking,
The mixture was filtered to remove a small amount of unreacted 7-amino-3-(3
-hydroxypyridazin-6-ylthiomethyl)-3-
Cefem-4-carboxylic acid was removed. 100 ml of acetonitrile was added to the filtrate, and the mixture was left at room temperature for 1 hour. The precipitate was collected by filtration, washed with 10 ml of water,
When dried under vacuum, 7-(0-aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid 0.49
7 (48%) were produced. Melting point 200-210°C (decomposed). Example 25 7-0-aminomethylphenylacetamide 3-(3-
Hydroxypyridazin-6-ylthiomethyl)-3-
Cefm-4-carboxylic acid 350 ml of dry tetrahydrofuran (THF') and N
Add four drops of -N-dimethylbenzylamine and powder well. Sodium O-[1-carbomethoxypropen-2-ylaminomethyl]phenylacetate 24. O7 (0.08
mol).

The suspension is stirred vigorously and cooled to -35-40° in an acetone-dry ice bath. A suspension of 13.67 (0.06 mol) of isobutyl hechloroformate is added all at once while keeping the temperature below -35°. After 2.5 minutes, the acetone-dry ice bath was removed and 0.7-amino-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid was added. 6 moles of O and N-methylmorpholine5. Add a solution of O4 (0.050 mol) to 250 ml of water.

The reaction mixture is stirred until it reaches room temperature (1-2 hours).
THF is removed under reduced pressure. 100 ethyl acetate on top of the aqueous solution
ml to form a layer, and add concentrated HCl'('pH 1.7-2
．． Set to 0. 3. Sprinkle the mixture with activated carbon (“Darco KB′”).
57 and filter. Separate the layers and extract the aqueous layer with 2 x 100 ml of ethyl acetate. The water phase was then filtered through filter paper and the pH was adjusted to 4. Adjust to O. After stirring, leave at 5-7° overnight.
Leave the fermented liquid to crystallize. The product is collected by filtration;
Wash with a small amount of water and dry in a P205 vacuum desiccator. 7-0-aminomethylphenylacetamide-3-
(3-hydroxy-pyridazin-6-ylthiomethyl)
The yield of -3-cephem-4-carboxylic acid is about 30%.

Example 26 7-(0-aminomethylphenylacetamide)-3-
(3-hydroxy-pyridazin-6-ylthiomethyl)
A suspension of the zwitterionic form of -3-cephem-4-carboxylic acid (0.361y) in 3 ml of methanol is cooled with ice and treated with a few drops of concentrated hydrochloric acid until a clear liquid is obtained. .

When ether is added, 7-(0-aminomethylphenylacetamide)-3-(3-hydroxy-pyridazine-6
A precipitate of -ylthiomethyl)-3-cephem-4-carboxylic hydrochloride salt forms as a light brown solid, which is filtered, collected and dried under vacuum over P2O5. Example 27 7-(0-aminomethylphenylacetamide)-3-
(3-hydroxy-pyridazine-6-ylthiomethyl)
To a stirred zwitterionic suspension of -3-cephem-4-carboxylic acid (0.361y) is added 1N sodium hydroxide solution at room temperature until a clear liquid (PHlO.8) is obtained.

When this solution is immediately lyophilized, an impure solid 7-(0-aminomethylphenylacetamide)-3-(3
-Hydroxy-pyridazine-6-ylthiomethyl)-3
- Cefem-4-sodium carboxylate is produced. Manufacturing method example 28 of dimethane sulfonated product Method: Jyoro i0-aminomethylphenylacetamide)
-3-(3-Hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid -2.0 mol of 4-carboxylic acid (approximately 980 f as anhydride) and 540 y of formaldehyde sodium sulfite (4.03 mol) and water 30
00 ml and 2700 ml (4.87 mol) of 30% SEH (sodium 2-ethylhexanate) acetone solution were placed in a suitable container and the mixture was heated at 40-45°C with stirring.
Heat to.

The mixture dissolves in about 10 minutes and becomes a yellow solution. heating 1
After 5 minutes, add 50y of decolorizing activated carbon (1 Dalco KB'') to the solution.
and further stirred at 40-45°C for 15 minutes.

After a heating reaction at 40-45° C. for a total of 30 minutes, filter with diatomaceous earth (1” dicalide).

Wash the activated carbon lump with 2000ml of 50% ethanol water. The filtrate was collected and adjusted to 25°C, and the solution was stirred at high speed at 25°C with 100% ethyl alcohol 1121? Add to.

7-(0-aminomethylphenylacetamide)-3-
(3-hydroxy-pyridazine-6-ylthiomethyl)
A pure white, amorphous precipitate containing two sodium methanesulfonate molecules attached to sodium -3-cefemu-4-carboxylate is formed.

The suspension was stirred for about 10 minutes and then, after filtration, the clumps were
Wash with 0% ethyl alcohol 151.

The mass is dried in a drier with air circulation at 50-55°C for about 2 hours and then under 4-6 vacuum for 24 hours.
What is obtained is 7-(0-aminomethylphenylacetamide)-3-(3-hydroxy-pyridazine-6-
It is an amorphous, white solid 1200-14007 consisting of two sodium methanesulfonate molecules attached to sodium ylthiomethyl)-3-cephemu-4-carboxylate.

The product generally contains several percent water and may contain a small amount of ethanol. This product is 71[0-
It is also named sodium N-N-bis(sodium sulfomethyl)aminomethylphenylacetamide]-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylate.

Example 29 Prepare the following mixture: Formaldehyde sodium sulfite 2.197 (2 equivalents) 71 (0-aminomethylphenylacetamide 3
-(3-Hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid zwitterion 3.5
7 (100-200 mesh) 25 ml of water (amount can be changed) 30% SEH-isopropanol solution 1
After stirring vigorously for about 0.5 hour at 4ml and 124°C, a nearly homogeneous solution is obtained.

Rapidly raise the temperature of the mixture to 40-43°C. This is allowed to stand for about 2 minutes and then immediately cooled to 20-23°C. Filter the solution to remove some insoluble material (do not stay in solution for more than 2 hours).

The solution at pH 7.3 is added over 5 minutes to 600 ml of very rapidly stirring pure ethanol (other alcohols such as anhydrous isopropanol may be used as alcohol). 7 An amorphous precipitate with two sodium methanesulfonates attached to sodium 7-(0-aminomethylphenylacetamide)-3-(3-hydroxy-pyridazin-16-ylthiomethyl)-13-cephem-4-carboxylate is formed. do.

Stir the mixture for 5 minutes. Filter and collect the precipitate,
Wash with 1 m of ethanol (or isopropanol) and vacuum dry at 50°C for 24 hours. The yield is about 4.370. The product has a pH of about 7 and is soluble in water to at least 200 m/ml.

Such solutions are stable at room temperature for at least 2 hours;
It is stable for even longer periods in dilute solutions. The product exhibits the same antimicrobial spectrum as the starting zwitterion and is fully biologically active whether or not it is hydrolyzed back to the zwitterion. Example 30 Sodium 7-[0-N-N-bis(sodium sulfomethyl)aminomethylphenylacetamide]-3-(3-hydroxy-pyridazin-16-ylthiomethyl)-3-cephem-4-carboxylate.

Production method 7-1 using hydroxymethane sulfonate (0-
Aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid zwitterion 1f7 (2.05 mmol) and sodium hydroxymethanesulfonate monohydrate 1.56y (10 mmol) and 6 ml of ethyl acetate solution (6 mmol) of 1 MSEH and 10 ml of isopropanol.
A mixture of 1 and 10 ml of water was stirred at room temperature for 3.5 hours.

The reaction solution was treated with 1V activated carbon, poured into 300ml of pure ethanol with stirring, and the mixture was stirred for 30 minutes at room temperature to obtain a crystalline product. Filter this,
Pure ethanol 50111 - 5 washes 3 times for 20 hours 4
It is well soluble in water (〉1y/ml) when dried over P2O5 under the conditions of 5 to 52 mm/l.
1.647 of sodium sulfomethyl)aminomethylphenylacetamide]-3-(3-hydroxy-pyridazin-16-ylthiomethyl)-13-cephem-4-carboxylate was obtained. The melting point is over 270 na. IR:ν
KBrl76Oll68O-1620, Maxl58O
ll4OOll2OOllO4O(1771-10uv
:λH2O249nm (ε14900)0m0ゝ272
nm (ε10900, Sh), 320nm (ε2100
、． sh).

NMR: δD2O3.3O~4.0 (6H,.m..S
-PpmCH2, φ-CH2-COlφ-CW2-N to 3-CH2-S), 4.04 (2H.S.N-CH
2φ to 3-CH2-S), 4.30 (41i.S,
N-CH2-SO3Na), 4.89 (1H.d, 4H
z16-H), 5.40(1H,.d14Hz17-H
), 6.78 (1H,.d19・5HZ・pyridazine-H), 7.1-7.4 (4H1 phenyl-H), 7.
32 (1H, .d19.5 ratio, pyridazine-H).

Example 31 Sodium 7-[0-N-N-bis(sodium sulfomethyl)aminomethylphenylacetamide]-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylate.

Manufacturing method using formalin and sodium sulfite (a) 3
0% formalin 1mt! 7-(0-aminomethylphenylacetamide)-3-(3
-hydroxypyridazin-6-ylthiomethyl)-3-
Zwitterion of cefem-4 carboxylic acid 0.975y(2
6 ml of 1M SEH solution and 10 ml of isopropanol were added.

The mixture was stirred at room temperature for 2.5 hours and 300 m
I poured it into 1. The produced sodium 7-[0-N-N-bis(sodium sulfomethyl)aminomethylphenylacetamide]-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cephem-4-carboxylate was filtered. It was collected, washed three times with 50 ml of ethanol each time, and dried under vacuum. The yield is 1.60y0(b) 71(0
-aminomethylphenylacetamide)-3-(3-hydroxypyridazin-6-ylthiomethyl)-3-cephem-4-carboxylic acid zwitterion 0.9757 (2 mmol), 6 ml of 1M SEH ethyl acetate solution, and 10 ml of isopropanol. 1 ml (10 mmol) of 30% formalin was added to a mixture of 10 ml of water.

The mixture was stirred at room temperature for 2 hours, resulting in a clear solution with a small amount of oily precipitate. After adding 1 (i!) of sodium sulfite, the solution was stirred for an additional 2 hours.

In the meantime, the oily precipitate had dissolved into solution. The reaction mixture was poured into 300 ml of ethanol with vigorous stirring to give 71[0-N-N-bis(sodium sulfomethyl)].
1.84y of sodium aminomethylphenylacetamide]-3-(3-hydroxy-pyridazin-6-ylthiomethyl)-3-cefemu-4-carboxylate was obtained,
This was collected by filtration, washed three times with 50 ml of ethanol, and dried under vacuum. Embodiments of the present invention and related matters are as follows.

1. The method according to claim 1.

2. The method according to claim 2.

Compounds of the following formula (where X is H or 0H and m is O or) and non-toxic pharmaceutically acceptable salts thereof.

6 Zwitterionic form of the compound of item 5.

7 Sodium and potassium of the compound of item 5.

However, when X is 0H, m is O. ) and nontoxic, pharmaceutically acceptable salts or easily hydrolyzable esters thereof. The following formula [where X is H or 0H. m is O or 1 (however,
is 0H, m is O)], non-toxic pharmaceutically acceptable salts thereof, and easily hydrolyzable esters thereof.

A compound represented by the following formula and a non-toxic, pharmaceutically acceptable salt thereof.

11 Zwitterionic form of the compound of item 10.

12 Sodium and potassium of the compound of item 10:
Mu salt.

13 Hydrochloride of the compound of item 10 14 below formula The compound shown in

15 below formula The compound shown in

16 Zwitterionic form of the compound of item 15.

17 Sodium and potassium salts of the compounds of paragraph 15.

18 Hydrochloride of the compound of item 15.

19 below formula The compound shown in

20 A compound represented by the following formula, a nontoxic pharmaceutically acceptable salt thereof, and an easily hydrolyzable ester thereof.

21 Sodium and Potassium Salts of the Compounds of Section 20.

22. The method of Section 1 described in Example 22.

23. The method of Section 2 described in Example 23.

24 The compound of item 3-21 produced by the method of item 1-2.

25. A pharmaceutical composition comprising a compound, salt or ester of paragraph 4-21 and one or more pharmaceutical carriers.

26. A method for the treatment of chronic diseases caused by Gram-positive and Gram-negative bacteria, which comprises administering an effective amount of the compound salt or ester of Items 4 to 19, or the composition of Item 25. .

27. A method for treating mastitis in cattle, comprising administering an effective amount of a compound, salt or ester of Items 4 to 19, or the composition of Item 31.

28 Pharmaceutical compositions described herein.

29. The method of items 26 and 27 described in the specification of the present invention.

Claims (1)

[Claims] 1. The following formula▲There are mathematical formulas, chemical formulas, tables, etc.▼ [Here, X is H or OH, and m is 0 or 1 (however, when X is OH, m is 0)] The following formula ▲Mathical formulas, chemical formulas, tables, etc. are included in the method for producing the compound represented by ▼ or its salt that is pharmaceutically acceptable, and its easily hydrolyzable ester ▼ Acylation of the acid whose ester is decomposed by the following formula ▲Mathematical formula, chemical formula, table, etc.▼III (where B is an amino protecting group, and the definitions of m and X are the same as the previous definition) By reacting with a derivative, the following formula ▲ There are mathematical formulas, chemical formulas, tables, etc. ▼ IV (Here, the definitions of B, X, and m are the same as the previous definitions.)
A compound of formula I or a salt thereof or an easily hydrolysable ester thereof is prepared, and then the amino protecting group is removed to form a compound of formula I or a pharmaceutically acceptable salt thereof or an easily hydrolysable ester thereof. A method comprising producing an ester.