JPS6160816B2 - - Google Patents

Description

[Detailed description of the invention] Thienamycin and its semisynthetic derivatives, epithi
Enamycin, a fused ring β-lactone containing olivanic acid
A new group of antibiotics has been described. Below
These compounds which will be defined in more detail below
These compounds are hereinafter referred to as “thienamycin compounds”.
It will be revealed. These compounds have high levels of antibacterial activity.
but undergoes extensive metabolism by mammals.
Ru. The primary metabolic site was confirmed to be the kidney.
Thienamycin by hydrolysis of β-lactam
An enzyme that catalyzes the inactivation of cancer is purified from kidney extracts.
has been done. Cytological localization, substrate specificity, and
Depending on indicators such as sensitivity to enzyme inhibitors,
This enzyme has been extensively studied in kidney dipeptidae.
dehydropase (E.C.3.4.13.11);
It is also called ``putidase,'' but it is not the same as ``putidase.''
It turns out that they are very similar, even if they are
Ru. However, β-lactamase activity
Shown only for syn-class compounds. In fact, β−
Penicillin, a representative member of the lactam antibiotic group
and via β-lactam cleavage of cephalosporins.
There is no precedent for mammalian metabolism to do so. The term “thienamycin compounds” refers to a common
naturally occurring with a fused β-lactam nucleus, or
is a semi-synthetic or synthetic derivative or analog composition.
It is used to secure members. These compounds
Products generally include 6- and (optionally) 2-substituted pens.
-2-m-3-carboxylic acid and 1-carbadecyl
Arpen-2-M-3-carboxylic acid, or 1-
Azabicyclo (3.2.0.) hept-2-ene-7-
It is classified as an on-2-carboxylic acid. Particular compounds particularly useful in the present invention are of the formula It is shown structurally in In the formula, X is CH₂or S
It is possible; R²is hydrogen; -S-
CH₂CH₂NHR³(R³is hydrogen, acetyl, formyl
midoyl, acetimidoyl);-S
(O)-CH=CHNHCOCH₃and -S-CH=
CHNHCOCH₃It is possible that R⁶teeth
[Formula] (R⁷is hydrogen, hydroxyl group or sulfony ) or R⁶is H. vinegar
All possible stereoisomers are included in the structure definition above.
included. All these compounds included in the formula are in the literature
It is written inside. X is CH₂,R²but
SCH₂CH₂N.H.₂,R⁶CH(OH)CH₃When
This compound is derived from S.cattleya (microorganisms).
Antibiotics produced by fermentation of Koken Bacteria No. 3309)
Biological agent thienamycin, issued on April 13, 1976.
Described and claimed in U.S. Pat. No. 3,950,357
has been done. N-substituted derivatives of thienamycin,
In other words, in the above formula, R³is other than hydrogen
is a co-pending U.S. patent application and its published
Disclosed and claimed in foreign equivalents
It is. Fermentation product N-A also called 924A
Cetylthienamycin (R⁶is CH(OH)CH₃And
R, R³is acetyl) was issued on May 16, 1977.
Claimed in Belgian Patent No. 848346 of
There is. N-imidoyl derivatives were issued on May 20, 1977.
Covered in Belgian Patent No. 848545 of
Ru. N-acetyl-dehydrothienamycin, or
is 924A_FiveCompounds containing unsaturated side chains called
is U.S. Patent Application No. 788491, filed April 18, 1977.
No. 1, Case 16022, and Base issued October 17, 1978.
The alcohol claimed in Lugie Patent No. 866035
It is a fermentation product. N-aceti also called 890A
luthienamycin, and 890A₃, desacetyl
890A, and desacetyl 890A₃The epibody of
French published patent application filed on November 19, 1976
No. 763887, US claiming priority of November 21, 1975
National Patent Application No. 634300 Case 15745, and 1977
Elucidated by Belgian Patent No. 848349 issued on May 16th
It is being done. 890A₂and 890A_FiveAlso called
The epi form of saturated thienamycin was published on April 28, 1976.
Claimed in French patent, Case 15839
ing. 890A₉or 890A_TenAlso called 6-sul
N-acetyl compounds containing fonyloxy are
French published patents filed on November 16, 1977
Application No. 7734456 (U.S. Patent No. 7734456, filed November 17, 1976)
Priority Case 15935) and November 16, 1977
French published patent application No. 7734456 (1976)
U.S. Patent Priority Claim, Case, November 17, 2017
15936). 890A₉as well as
890A_TenThe desacetyl analogs of
U.S. Patent Application No. 767723 filed on February 11th
15975, now abandoned and filed December 15, 1977 U.S.
Patent Continuation Application No. 860665 and February 9, 1978
French Patent Application No. 7803666; 1977 2
U.S. Patent Application No. 767920, filed May 11, Case
15976, now abandoned and filed on January 25, 1979.
National Patent Continuation Application No. 006959 and February 9, 1978
No. 7,803,667 filed in French Patent Application No. 7803667
being sought after. 890A₉and 890A_TenThis in the series
Some of the latter compounds are olibanic acid.
(Corbett et al. J.Chem.Soc.Chem.Commun.1977, 24
(see pages 953-954).
Ru. In the above formula, R²is hydrogen, des
Also called cysteaminyl thienamycin
is a U.S. patent application filed on March 22, 1976.
No. 668898, Case 15866, Now Abandoned, &
Continuing: U.S. Patent Application No. 31, 1977, filed October 31, 1977
No. 847297, now abandoned and issued November 20, 1978.
Claimed in Belgian Patent No. 867227
Ru. R⁶is hydrogen and X is CH₂These compounds are 1977
U.S. Patent Application No. 843171 filed on January 1, 2017
15902 and corresponding German published patent no.
2751624.1 (filed on November 18, 1977)
It is. R²ga-SCH₂CH₂R at NHAC⁶is C₂H_FiveChie who is
Namycin-type antibiotics are named PS-5 and K.
By OKaimura et al.J. Antibiotics31 volumes, 480 pages
(1978). About this
See also Rugie Patent No. 865578. The compound where X is S, also called “penem”, is R.
B. Woodward, “Chemistry of β-Lactam Antibiotics”
Recent Advances” J.EIKS (editor), British Chemical Society,
London, p. 167; R.B. Woodward, Pharmaceutical Research.
Recent topics: 500th anniversary of Uppsala University
Abstracts of the Posium, Uppsala, Sweden 1921
Page, October 1977,Acta.Pharm.Suecica, 14 volumes,
Supplementary page 23 and U.S. Patent No. 24, January 24, 1978.
Described in issue 4070477. Particularly preferred composition in thienamycin compounds
The members are N-formimidoyl and thienamycin.
It is an N-acetamidoyl derivative. recently described
of N-formimidoylthienamycin, which is
Crystalline forms are also useful in the practice of this invention.
A working example illustrating a preferred method of making this compound.
One example is as follows. Illustrative examples N-formimidoyl thienamycin, crystals Step A Benzylformimide, hydrochloride Separate addition funnel, overhead stirrer,
and 3 three-necked flasks equipped with a reflux condenser.
Benzyl alcohol (125g, 1.15mol), form
Amide (51 g, 1.12 mol) and anhydrous ether
(1200ml). This mixture in a nitrogen atmosphere
Stir vigorously at room temperature (20-25℃) and add 50 ml of anhydrous ether.
Benzoyl chloride (157 g, 1.12 moles) in
Add dropwise using addition funnel. Approximately 50 minutes for addition
It takes. The reaction mixture is stirred for a further 60 minutes at room temperature. de
Remove the ether using Kant's method and add 500 ml of anhydrous ether.
Add 300 ml of acetic anhydride in solution. this mixture
Stir for 30 minutes at room temperature. Let the precipitate sink and deca
The ether-acetic anhydride is removed again by the solvent method. solid
Collect the mold by filtering and wash with 500ml of ether.
When KOH is added in vacuum and dried for 2 hours at 25℃, 130
g (67%) of benzylformimidate hydrochloride
Obtained as a white solid. The NMR of this product is δ(DMSO)5.7(S,
2H, φCH₂), 7.5 (S, 5H, φ), 9.0 (S,
1H, HC=N). This product is thermally unstable
It is. At 0°C and above, this compound
decomposes into lumamide and benzyl chloride. but,
After storage at -20℃ for 2 months, there is no appreciable decomposition.
It was not recognized. Step B Derivatization of Thienamycin Thienamycin (concentrate from fermentation process, 28
Contains g of thienamycin. Aqueous solution of 6 at PH6.5
liquid) in a large beaker (12).
and cool to 0°C. This beaker has a PH meter.
- Equipped with an efficient high-speed stirrer.
Adjust the pH to 8.5 by carefully adding 3N KOH
(Drop KOH into the stirring liquid with a syringe.)
). Add this solution to 6 equivalents of solid benzyl fluoride.
Treated in small portions with muimidate hydrochloride (~100g)
do. During this time, add 3N KOH (200ml) using a syringe.
The pH is maintained at 8.5±0.3 by adding Attachment
Addition takes 3-5 minutes. The reaction mixture was stirred at 0 °C for 6 min, then the reaction
Liquid chromatography to ensure completion of
Test with. Adjust the pH of this solution to 7 with 1NHCl.
Ru. Measure the volume of the reaction mixture and expose the solution to UV light.
More testing. Neutralized reaction mixture at <10°C
Concentrate to 15 g/ml using a reverse osmosis unit. This dark
Measure the volume of condensate and adjust the pH to 7.2-7.4 if necessary.
Adjust. Pour this concentrate into a medium pore glass sieve.
filter to remove solids present after concentration.
Ku. Stage C Dowex 50W x 2 Chromatograph
E This concentrate (750-1000ml, 15-20g)
50W x 2 (200-400 meters)
18 columns of pre-cooled resin (resin) at 0°C.
Load the column with distilled, deionized water to increase the flow rate.
90ml/min, 0-5 at 0-45psig pressure above column
Elute at ℃. Collect the first categories 4, 2, and 1 that appear.
Then, collect 18 sections of 450ml each, and finally
Collect the 2 categories. Measure the UV of each category
(1/100 dilution, NH₂(Excluding absorption by OH)
Calculate the total amount of NFTs present in each segment. most
The first and last divisions are made by liquid chromatography.
Verify the purity and match the desired classification with large amounts of ingredients.
Ru. The PH of the categories with large amounts of components is PH makeup.
Determined by tar and bromothymol blue indicator reagent.
If necessary, adjust the pH to 7.2-7.4. The ingredients are
The bulk of the combined fractions (3-4) were then exposed to UV.
Determine the total formamidine content. 15
-16 g and the yield from the column is 75%. child
The fraction containing a large amount of components was subjected to reverse osmotic pressure at <10°C.
Concentrate as much as possible in the unit and then use a rotary evaporator.
33g/at below 28℃ using a rotor
Concentrate. A total liquid volume of approximately 500 ml of concentrate is obtained. Step D N-formimidoyl thienamycin
crystallization of Adjust the pre-stage concentrate to pH 7.3 if necessary,
UV N-formimidoylthienamycin content
It is about 85-90% when quantified by . Gather this concentrate.
With a lath filter (one with medium pores)
Drain and collect in a large Erlenmeyer flask. 3A Etano
into this concentrate by straining 5 volumes (2200 ml) of the
and incubate the solution for 10 min at room temperature and at 0 °C for 12-24 min.
Stir for an hour. The crystals were filtered by suction at 80% 3A at 0℃.
with 0.1 volume (~250 ml) of ethanol, then at room temperature.
Wash with 1/25 volume (100 ml) of 3A ethanol. crystal
When dried in vacuum for 12-24 hours, the total yield is about 40%.
-formimidoylthienamycin is obtained
(10-12g). N-formimidoylthie prepared as above
The analysis results for Namycin 50g are as follows.
be. C, theoretical value 45.42%, actual value 45.82% H, theoretical value 6.03%, actual value 5.72% N, theoretical value 13.24%, measured value 13.10% S, theoretical value 10.10%, measured value 10.14% Predicted value of residue when burned: 0.5, actual value: 0.47
%, [α]²⁵ _D=89.4゜, T.G.=6.8%, UV λmax
300nm, E¹%=328. Dipeptidase inhibitor Select the metabolism of dipeptidase [E.C.3.4.13.11].
Chemicals that selectively inhibit dipeptidases, also known as “dipeptidases”
“inhibitor” has the following formula Z-2-acylamino-3-mono-substituted protein having
Includes the chemical compound that is ropenoate. During the ceremony
R²and R³are in the range of 3-10 and 1-15 carbon atoms, respectively.
These are the hydrocarbon groups in the circle. These hydrocarbon groups R²Reach
BiR³Up to 6 hydrogens on either side of
may be substituted with a halogen, or a non-terminal metal may be substituted with a halogen.
Oxygen or sulfur (including this oxidized form)
) may be replaced. R³The terminal hydrogen of
for example, by alkanoyl acids of 1-8 carbon atoms.
acylation, or alkyl and dialkyl groups.
May be carbamoylated including bamate derivatives
), or hydrogen can be replaced with a
mino group (this is acylamino, ureido, amidi
guanidino or quaternary nitrogen group-containing alkyl or
Even if it is a derivative of a substituted alkylamino group,
(good) or alternatively
carboxylic acid, phosphoric acid, sulfate group, or their
With acid groups such as esters, amides, and cyano groups
Substitutions are also possible, or the terminal amino acid group
It can also be a combination of them, such as
stomach. R²is a branched alkyl or cycloalkyl group (C₃
−C_Ten), but next to the carbonyl group
The adjacent carbons must not be quaternary. R¹is water
elementary, lower alkyl (C_1-6) or dialkylamino
Alkyl [e.g. -CH₂CH₂N(C₂H_Five)₂,−
CH₂CH(CH₃)N(CH₃)₂]. Some of the compounds of the above formula are asymmetric
Has a type. Racemic Z-2-(2,2-dimethy
cyclopropanecarboxamide)-2-octane
Non-acids are optically resolved. Activity is dextrorotatory
isomer, which has the S-coordination. R²The definition includes the following subgroups: −R^Four A R in the formula^Fouris a straight chain, branched chain or chain of 3-10 carbon atoms.
is a cyclic hydrocarbon group, and R²with the definition of
May be substituted as specified, −R^FiveR⁶ B R in the formula^Fiveis cycloalkyl of 3-6 carbon atoms
Yes, R⁶is a 1 or 2 alkyl substituent (these
Another ring is spliced onto the cycloalkyl group.
) or R^FiveReach
BiR⁶is the above-mentioned R²Replaced as specified in the definition of
It is okay to be −R⁷R⁸ C R in the formula⁷is an alkylene group with 1-3 carbon atoms
Yes, R⁸is cycloalkyl of 3-6 carbon atoms
However, these are the R²and R⁶The definition of
May be replaced as described, Within these subgroups are the following specific compounds:
Ru, A: Z-2-isovaleramide-2-pente
Noic acid; methyl Z-2-isovaleramide-2
-butenoate; Z-2-isovaleramide-2
-butenoic acid; Z-2-benzamido-2-butenoic acid;
Thenoic acid; Z-2-(3,5,5-trimethyl
hexaamide)-2-butenoic acid;
Clobutanecarboxamide-2-butenoic acid;
Z-2-cyclopropanecarboxamide-2-b
Thenoic acid; Z-2-cyclopropanecarboxa
Mido-2-pentenoic acid; Z-2-(3-methyl
ruberalamide)-2-butenoic acid; Z-2-
Cycloheptanecarboxamide-2-butenoin
Acid; Z-2-nonanamido-2-butenoic acid;
Z-2-cyclohexanecarboxamide-2-bu
Thenoic acid; Z-2-(4-methylvaleramide
Z-2-t-butenoic acid;
Cetamido-2-butenoic acid; Z-2-octa
Amido-2-butenoic acid; Z-2-butylamide
Do-2-butenoic acid; Z-2-valeramide-
2-butenoic acid; Z-2-valeramide-2-
Pentenoic acid; Z-2-cyclopentanecarbo
Xamido-2-butenoic acid; Z-2-(6-methane)
tilheptaamide)-2-butenoic acid; Z-2
-Hexamido-2-butenoic acid; Z-2-
(3,7-dimethyloctanamide)-2-buteno
Inic acid; Z-2-(3,7-dimethyl-6-oc
Z-2-(5-
chlorovaleramide)-2-butenoic acid; Z-
2-(3-chlorobenzoylamide)-2-buteno
Inic acid; Z-2-(2-chlorobenzamide)-2
-butenoic acid; Z-2-nonamido-2-butenoic acid
Noinic acid; Z-2-(6-bromohexanamide
Z-2-(3,3-dimethyl)-2-butenoic acid;
tinpropenamide)-2-butenoic acid; Z-
2-Benzamido-2-cinnamic acid; Z-2-benz
Amido-2-pentenoic acid; Z-2-benza
Mido-5-methoxy-2-pentenoic acid; Z-
2-Benzamido-2-hexenedionic acid; Z-
2-isovaleramide-2-octenoic acid; Z
-2-isovaleramide-2-cinnamic acid; Z-2-
Isovaleramide-2-hexenedionic acid; Z-
2-cyclopropanecarboxamide-2-cinnamon
Acid; Z-2-cyclopropanecarboxamide-2
-hexenedionic acid; Z-2-(5-methoxy-
3-methylvaleramide)-2-butenoic acid;
Z-2-ethylthioacetamido-2-butenoy
acid; Z-2-(2,2-dichlorocyclopropane)
carboxamide)-2-butenoic acid; Z-2
-(2-ethylhexanamide)-2-butenoin
Acid; Z-2-di-n-propylacetamide-2
-butenoic acid; B: Z-2-(2,2-dimethylcyclopropyl)
carboxamide)-2-butenoic acid; (+)-
Z-2-(2,2-dimethylcyclopropanal)
boxamido)-2-butenoic acid; Z-2-(2,
2-dimethylcyclopropanecarboxamide)-
2-pentenoic acid; Z-2-(2,2-dimethyl
cyclopropanecarboxamide)-2-octane
Noinic acid; Z-2-(2,2-dimethylcyclopropylene)
lopanecarboxamide)-2-hexenoic acid;
Z-2-(2,2-dimethylcyclopropanal)
boxamide)-2-cinnamic acid; Z-2-(2,2-di
Methylcyclopropanecarboxamide)-5-methane
Toxy-2-pentenoic acid; Z-2-(2,2
-dimethylcyclopropanecarboxamide)-
4,4,4-trifluoro-2-butenoic acid;
Z-2-(2,2-dimethylcyclopropanal)
boxamide)-3-(2-chlorophenyl)prope
Noinic acid; Z-2-(2,2-dimethylcyclopropylene)
lopanecarboxamide)-2-hexenedioin
Acid; Z-2-(2-ethylcyclopropanecarbo
xamido)-2-butenoic acid; Z-2-(2,2
-diethylcyclopropanecarboxamide)-2
-butenoic acid; Z-2-(2,2-diethylsiloxane)
(chloropropanecarboxamide)-2-pentenoy
Z-2-(2-isopropyl-2-methyl
cyclopropanecarboxamide)-2-butenoy
Z-2-(2-methylcyclohexanecal)
boxamide)-2-butenoic acid; Z-5-sia
No-2-(2,2-dimethylcyclopropanecar)
boxamide)-2-pentenoic acid; Z-5-
(N,N-dimethylcarbamoyl)-2-(2,2
-dimethylcyclopropanecarboxamide)-2
-Pentenoic acid; Z-2-(2,2-dimethyl
cyclopropanecarboxamide)-5-methane
Ruphonyl-2-pentenoic acid; Z-2-(2,
2-dimethylcyclopropanecarboxamide)-
5-ethoxycarbonyl-2-pentenoic acid;
Z-2-(2,2-methylcyclopropanecarbo
(xamide)-2-butenoic acid: methyl Z-2-
(2,2-dimethylcyclopropanecarboxamide
)-2-butenoate; ethyl Z-2-(2,2
-dimethylcyclopropanecarboxamide)-2
-butenoate; Z-2-(2,2-dimethylsiloxane)
(chloropropanecarboxamide)-2-butenoin
Acid 2-dimethylaminoethyl ester; Z-2-
(2,2-dimethylcyclopropanecarboxamide
d)-2-pentenoic acid 3-diethylaminope
Lopylester; Z-2-(2,3-dimethylsiloxane)
(chloropropanecarboxamide)-2-butenoin
Acid; Z-2-(3,3-dimethylcyclobutanka)
ruboxamide)-2-butenoic acid; Z-2-(2
-spirocyclopentanecarboxamide)-2-
Butenoic acid; Z-2-(2-t-butyl-3,
3-dimethylcyclopropanecarboxamide)-
2-butenoic acid; Z-2-(2,2-dimethyl
cyclopropanecarboxamide)-4-methyl-
2-pentenoic acid; Z-2-(2-t-butyl
cyclopropanecarboxamide)-2-butenoy
Z-2-(2-phenylcyclopropanka)
ruboxamide)-2-butenoic acid;
Chlohexyl-2-(2,2-dimethylcyclopropylene)
lopanecarboxamide)-propenoic acid; Z-
5-carboxy-5-(2,2-dimethylcyclo
propanecarboxamide)-4-pentanamide
Z-5-dimethylamino-2-(2,2-di
Methylcyclopropanecarboxamide)-2-pe
Ntenoic acid; Z-3-cyclopropyl-2-
(2,2-dimethylcyclopropanecarboxamide
d) propenoic acid; Z-2-(2,2-dimethyl
cyclopropanecarboxamide)-2,5-he
Xadienoic acid; Z-2-(2,2-dimethylsiloxane)
(chloropropanecarboxamide)-4-phenyl-
2-butenoic acid; Z-2-(2,2-dimethyl
cyclopropanecarboxamide)-6-mercap
Th-2-hexenoic acid; Z-2-(2,2-di
Methylcyclopropanecarboxamide)-5-methane
Chilthio-2-pentenoic acid; Z-2-(2,
2-dimethylcyclopropanecarboxamide)-
5-phosphono-2-pentenoic acid; Z-2-
(2,2-dimethylcyclopropanecarboxamide
Z-2-(2,2-di)-2-heptenoic acid;
Methylcyclopropanecarboxamide)-5-ph
Enyl-2-pentenoic acid; Z-2-(2,2
-dimethylcyclopropanecarboxamide)-2
-nonenoic acid; Z-2-(2,2-dimethylsiloxane)
(chloropropanecarboxamide)-2-decenoin
Acid; Z-2-(2,2-dimethylcyclopropane
carboxamide)-2-tridecenoic acid; Z-
2-(2,2-dimethylcyclopropane carboxy
(samido)-6-methoxy-2-hexenoic acid
(and 5-methoxy-2-pentenoic acid); Z
-2-(2,2-dimethylcyclopropanecarbo
xamido)-6-methyl-2-heptenoic acid;
Z-4-cyclohexyl-2-(2,2-dimethy
cyclopropanecarboxamide)-2-buteno
Inic acid; C: Z-2-cyclobutylacetamide-2-
Butenoic acid; Z-2-cyclopentylacetoacetate
Mido-2-butenoic acid; Z-2-cyclohexy
Ruacetamido-2-butenoic acid; Z-2-
(4-cyclohexylbutyramide)-2-buteno
Inic acid; Z-2-cyclopropylacetamide-
2-butenoic acid; Z-2-cyclopropyl acetate
Toamido-2-pentenoic acid; Z-2-(3-
cyclopentylpropionamide)-2-buteno
Inic acid; Z-2-(3-cyclohexylpropio
Z-2-(4-
(2-thienyl)-butyramido)-2-butenoy
Z-2-(4-phenylbutyramide)-2
-butenoic acid; Z-2-(D,L-α-lipore
mido)-2-pentenoic acid; Z-2-(D,L-
α-lipoamide)-2-cinnamic acid; Z-2-(3-
(2-tetrahydrofuryl)-propionamide)
-2-butenoic acid. R mentioned above²Particularly preferred substituents in the definition of
are 2,2-dimethylcyclopropyl and 2,2-
Contains dichlorocyclopropyl group. R³A particularly preferred group of compounds in the definition of
Quaternary nitrogen, amine derivative or amino acid derivative group
N-alkyl with certain terminal substituents (1-9 carbons)
(1-9 carbons) and N-methyl (1-9 carbons). The term “quaternary nitrogen” refers to positively charged tetrasubstituted or
Refers to multiple aromatic ring nitrogens. same or different1
-substituted with a hydrocarbon substituent having 7 carbon atoms
It also means an ammonium group moiety. The term “amino derivative” refers to amino, acylami
No, ureido, amidino, guanidino and the like
means an alkyl derivative of The term "amino acid derivative group" refers to cysteinyl (-
SCH₂CH(NH₂) COOH) Sarcosyl (-N
(CH₃)CH₂means a group such as COOH). child
In the case of bonding with O, N, or S of known amino acids
The hydrogen present is replaced. R²and in the most preferred group having R substituents
Particularly preferred compounds are R²is 2,2-dimethy
cyclopropyl or 2,2-dichlorocyclopropyl
Lopil and R³has no terminal substituents. Some
is trimethylammonium, amidino, guanidi
-, 2-amino-2-carboxyethylthio, or
is 3 carbon atoms with a terminal substituent that is ureido
to 7 hydrocarbon chains. these
Specific example names include: Z-2-(2,2-dimethylcyclopropanka)
ruboxamide)-8-trimethylammonium
Hydroxide-2-octenoic acid inner salt; Z-2-(2,2-dichlorocyclopropanka)
ruboxamide)-8-trimethylammonium
Droxide-2-octenoic acid inner salt; Z-2-(2,2-dimethylcyclopropanka)
ruboxamide)-8-amidino-2-octenoy
acid; Z-2-(2,2-dimethylcyclopropanka)
ruboxamide)-8-guanidino-2-octeno
inic acid ;Z-2-(2,2-dimethylcyclopropane
carboxamide)-8-ureido-2-octeno
Inic acid; Z-8-(L-2-amino-2-carboxye
tylthio)-2-(2,2-dimethylcyclopropyl)
carboxamide)-2-octenoic acid; Z-2-(2,2-dimethylcyclopropanka)
ruboxamide)-2-octenoic acid (racemic
and dextrorotator); and Z-2-(2,2-dichlorocyclopropyl)
carboxamide)-2-octenoic acid. Z-coordination (J.E.Blackwood et al., J.Amer.Chem.
Soe. vol. 90, p. 509 (1968)) by A. Srinavasan et al.
The work of [Tetrahedron Lett., p. 891 (1976)]
Similarly, for the above compounds, the NMR spectra
Attribution was made based on the file. R¹These compounds of the formula where is H are free acids
Although it is described and named as
For those in the know, alkalis and alkaline
Various lithium metal, ammonium, or amine salts, etc.
Pharmaceutically acceptable derivatives of are also equivalent to the free acid.
It is clear that it is possible to adopt
Will. Sodium, potassium, calcium or
Salts such as tetramethylammonium salt are suitable for
ing. Some of the compounds of formula are new compounds.
Yes, pending U.S. patent application filed July 24, 1978.
Application No. SN972212, current… and concurrent with this patent
U.S. Patent No. SN filed in
(which does not form part of the invention)
ing. As mentioned above, thienamycin-type compounds
Used in combination with putidase inhibitors. METHODS OF TESTING AND USING THE INVENTION As mentioned above, thienamycin, its natural derivative
By studying the properties of conjugates and their semi-synthetic derivatives.
The various species tested (hatsuka rat, mouse,
in dogs, chimpanzees, laser monkeys)
Main surrogate decomposition routes for removing these compounds
was revealed. The degree of metabolism is low
This is reflected in the yield and short half-life of blood. this minute
The properties of the solution were first determined by Bergman, M. and
Schleich, H. Z.Physiol.Chem. vol. 205, p. 65
Kidney dipeptidase described in (1932)
(E.C.3.4.13.11) is the cleavage of the lactam ring by
It has been shown that Further Greenstein, J.P.
Advance in Enzymology, Vol., Wiley-
Interscience (1948), New York and
Campbell, B.J.; Lin, Y-C., Davis, R.V. and
and Ballew, E., “Purification of Granular Kidney Dipeptidase.”
and properties”, Biochim.Biophys.Acta, volume 118, p. 371
(1966). Formula compounds that inhibit the action of renal dipeptidase
Added an in vitro search method to demonstrate the ability of
reached. This is dipeptine isolated from pig kidneys.
Glycyldehyde by solubilized preparation of Dase
Inhibits hydrolysis of lophenylalanine (GDP)
It measures the ability of a compound to Operability
is as follows: 50mM “MOPS” (3-(N
-morpholino)propanesulfonic acid) buffer, PH
7.1 in a 1 ml system containing 5 μg of lyophilized
Add enzyme and test compound to a final concentration of 0.1mM.
Add to sea urchin. After 5 minutes at 37°C, remove the GDF.
Add so that the final concentration is 0.05mM. 37 more
Keep at 275 nm absorption for 10 min at 275 nm for GDP hydrolysis.
It is measured by the change over time. Enzyme inhibition is
Measured against standards tested without harmful agents,
Expressed as inhibitor binding constant Ki. It contains 50% enzyme
is the concentration of inhibitor that inhibits. This search method replaces the substrate GDP with thienamycin.
Adopt with priority. Because GDP is a kidney disease
Has a higher maximum rate of hydrolysis by putidase
and is thereby required. reduce the amount of enzyme
This is because it can be done. GDP and Chinama
Both icins are similar parents to renal dipeptidases.
Furthermore, the Ki of the tested inhibitor was
was the same for the two substrates. In addition to this in vitro search method,
By increasing the urinary recovery of thienamycin
Measuring the ability of a test compound to inhibit metabolism as reflected
In vivo tests were also conducted. This method of operation requires static
10-100mg/Kg of test compound via intravenous or subcutaneous route
together with thienamycin at a dosage rate of 10mg/Kg.
to be administered. Thie in the urine for a period of then 4 hours
The recovered amount of namycin was administered together with the test compound.
Compare with the amount recovered in the standard group. The amount of thienamycin recovered in urine was
If applicable, those described in U.S. Patent No. 3,950,357
Measured by cylinder or disk diffusion method
Ta. thisThe bioassay method using Staphylococcus aureus ATCC6538 as the test bacterium ranges from 0.04 μg/ml to 3.0 μg/ml.
Shows useful correspondence in the ml range. The combination of an inhibitor and a thienamycin class compound can be formulated into a pharmaceutical composition containing the two compounds in a pharmaceutically acceptable carrier. The two compounds have a weight ratio of thienamycin compound to inhibitor of 1:3 to 30:
1, and preferably from 1:1 to 5:1. These components can also be administered separately.
For example, administer thienamycin compounds intramuscularly or intravenously at 1-100 mg/Kg/day, preferably 1-20 mg/day.
Kg/day or 1-5 mg/Kg/day in divided doses (for example 3 to 4 times per day). The inhibitor may be administered orally, intramuscularly, or intravenously at 1-100 mg/Kg/separately from the thienamycin compound.
day, also preferably 1-30 mg/Kg/day, or 1-5
Amounts of mg/Kg/day can be administered. It is desirable that the amounts of the two components administered during the day be within the proportion limits set forth above. A sterile intravenous aqueous solution of two crystalline compounds, one N-formimidoylthienamycin and the other (+)Z-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid. When administered as a single dose (sodium salt), the currently known most preferred dosage for patients is 150 mg of thienamycin;
Octenoic acid is 75 or 150 mg. This dosage is administered to humans (assuming their body weight is about 80 kg) 1 to 4 times per day, or 2-8 mg/Kg/day of the thienamycin compound and 1-4 mg/kg/day of the inhibitor.
Administer 8mg/Kg/day. These ingredients, whether administered separately or together, may be prepared in a pharmaceutically acceptable form, such as a conventional base suitable for oral administration, such as a capsule, tablet or liquid solution or suspension. used in a carrier.
The components can be dissolved separately or together in a suitable base for injectable administration. Compositions suitable for oral use include diluents, granulating agents, preservatives, thickening agents, flavoring agents, and coating agents. Examples of oral compositions with active ingredients or with the acid ingredient alone are prepared by mixing the dry powder with gelatin, starch, magnesium stearate, and alginic acid and forming tablets. As mentioned above, the currently known preferred method is parenteral administration of the thienamycin class compound and simultaneous parenteral administration or oral administration of the inhibitor compound. Examples illustrating the invention are as follows. Section 1 Examples Illustrating Activity Example 1 In Vitro Test Data A 1 ml system with 50 mM "MOPS" buffer, PH 7.1 is used. To this are added 5 μg of pig kidney enzyme and the test compound to a final concentration of 0.1 mM. After 5 minutes at 37°C, GDP is added to a final concentration of 0.05mM. The system is again kept at 37°C for 10 minutes. GDP hydrolysis at 275nm
It is measured by the change in optical absorption over time. Inhibition of this enzyme is determined in comparison to a standard similarly performed without inhibitor and is expressed as % inhibition. Ki is a constant indicating the concentration of inhibitor required to cause 50% inhibition of the enzyme. The value was calculated from several in vitro assays at inhibitor concentrations that resulted in inhibition of less than and greater than 50%. [Table] [Table] [Table] [Table] [Table] [Table] [Table] Example 2 In-vivo test data The in-vivo assay in rats is carried out as follows: 20 g of Charles River CD, Female rats are injected subcutaneously with the selected dosage of the chemical inhibitor. Approximately 2 minutes later, the dose of thienamycin is injected intravenously. The thienamycin standard described above is also run. The level of thienamycin in the urine as a % of the dose is determined by bioassay. The results are shown in the table. The numbers for the two test compounds are from the table. Compound 7 is 2-isovaleramido-2-butenoic acid and compound 10 is Z-2-cyclopropanecarboxamide-2-butenoic acid. [Table] Example 3 Compound 2-isovaleramido-2-butenoic acid, compound 7, and Z-2-(2,2-dimethylcyclopropanecarboxamide)-2-butenoic acid in combination with thienamycin (THM) A detailed study was carried out using in vivo tests using rats. The general operating method is the same as in Example 2. The results are summarized in tables and tables. Example 4 In another study in rats, the systemic antibacterial activity of thienamycin was compared to 2-isovaleramide-2.
- Increased approximately 3 times by co-administration of butenoic acid. See table. Table: Co-administered 2-isovaleramide on the systemic efficacy of thienamycin in the treatment of Staphylococcus aureus infections.
Effects of 2-butenoic acid [Table] [Table] (b) Simultaneous administration [Table] (b) Simultaneous administration.
Example 5 Male beagle dogs were used to study the effect of dipeptidase inhibitors on the urinary recovery of N-formimidoyl thienamycin. In a control experiment, the dog received 5 mg/Kg of N-formimidoylthienamycin intravenously without any inhibitor. In the second experiment, the same amount of N-formimidoylthienamycin was used, and Z-2-isovaleramido-2-butenoic acid was administered in three doses (20 mg/Kg of compound each time). The first dose was administered immediately after the N-formimidoylthienamycin injection, the second dose 40 minutes later, and the third dose 60 minutes later. In the third experiment, Z-2-
(2,2-dimethylcyclopropanecarboxamide)-2-butenoic acid was administered at once (2 mg/
Kg). The results are shown in the table. Table 2. Examples Illustrating Chemical Preparations The inhibitor compounds are novel compounds claimed in pending patent applications. These compounds are made by direct condensation of the appropriate 2-keto acid and amide; In the formula, R ² and R ³ are as defined. Typical reaction conditions include mixing the acid and amide in an inert solvent such as toluene or methyl isovalerate at about 1-
Mixing in a ratio of 4:1 and heating to reflux for 3-48 hours, preferably 5-24 hours with azeotropic removal of water. When cooled, the product usually precipitates out as crystals from the solution, but the product can also be isolated by base extraction. The product can be recrystallized using commonly known methods. In an optional variation of this method,
It is required to add a small amount of P-toluenesulfonic acid as a catalyst during the reaction. Another route to new inhibitor compounds uses t-butyl esters of alpha-amino acids in reaction with acid chlorides. This reaction takes place in a solvent such as methylene chloride in the presence of a base such as triethylamine. Generate N-
The acyl product () is then oxidized by treatment with t-butyl hypochlorite and then sodium methoxide is added. Thereby, the 2-methoxy derivative () and/or its elimination product, α,β-
An unsaturated ester () is obtained. Further treatment with anhydrous hydrochloric acid or (or a mixture of both) converts it to the desired α,β-unsaturated free acid (). [Formula] [Formula] [Formula] Certain compounds in which R ₃ has a terminal substituent that is an amino, quaternary nitrogen, thiol or carboxy derivative are intermediates having a brominated substituent in place of the desired terminal substitution. from, the desired thiol,
It can be most conveniently prepared by condensation with amino acids, acids, and the like. More details regarding the preparation of the compounds can be found in the Examples below. Example 6 1.07 g (10.5 mmol) of 2-ketobutyric acid and 0.71
A solution of g (7.0 mmol) of isovaleramide in 15 ml of toluene is stirred at reflux and the water is collected in a small Dean-Stark trap. After 5 hours, the solution is cooled and considerable crystallization occurs. After standing, collect the solids on a filter,
Wash _with toluene then _CH2Cl2 . Yield of white crystals = 0.47 g, melting point 172-174° (slightly before softening). This material is recrystallized from diisopropyl ketone. TLC (4:1 toluene-AcOH)
indicates that there are only trace amounts of other isomers. Yield of white crystals = 0.32 g (25%), melting point 175° (slightly softens before that). NMR showed essentially only the Z-isomer. Analysis (C ₉ H ₁₅ NO ₃ ) Calculated Value Actual Value C 58.36 58.59 H 8.16 8.55 N 7.56 7.43 Example 7 Z-2-(2,2-dimethylcyclopropanecarboxamide)-2-pentenoic acid 1.74 in 20 ml of toluene g (15 mmol) of 2
- A solution of ketovaleric acid and 1.13 g (10 mmol) of 2,2-dimethylcyclopropanecarboxamide is stirred and the water is collected in a small Dean-Stark trap. After 20 hours, the solution is cooled and treated with a gentle stream of _N2 . Before most of the solvent has evaporated, the walls of the container are scratched to cause crystallization. After standing, the solids are collected on a filter and washed with toluene and some _Et2O . Yield of white crystals = 0.63g (30%), melting point
154.5−155.5° (slightly softens before that).
TLC (4:1 toluene:AcOH) shows very little presence of other isomers. NMR is consistent with Z-configuration. Analysis (C ₁₁ H ₁₇ NO ₃ ) Calculated value Actual value C 62.53 62.86 H 8.11 8.27 N 6.63 6.75 Example 8 Z-2-(3-cyclopentylpropionamide)-2-butenoic acid 1.41 g (10 mmol) of 3- A solution of cyclopentylpropionamide and 1.53 g (15 mmol) of 2-ketobutyric acid is stirred and refluxed under a small Dean-Stark trap. After 8 hours, the solution is cooled and a large amount of crystals form. The solid is collected on a filter and washed with toluene and CH ₂ Cl ₂ . Yield of white crystals = 1.44 g, melting point 180.5-182° (pre-softened). Recrystallize this material from methyl ethyl ketone, yield of white needles = 0.63 g (28%),
Melting point 184-185° (slightly pre-softened). TLC
(4:1 toluene:AcOH) shows a single spot and NMR indicates substantially pure Z-isomer. Analysis (C ₁₂ H ₁₉ NO ₃ ) Calculated value Actual value C 63.97 63.99 H 8.50 8.67 N 6.22 6.27 Example 9 Z-2-(2-ethylhexanamido)-2-butenoic acid, 10 g of 2-ethylhexanoyl chloride is added to 25 ml of cooled concentrated NH ₄ OH solution with stirring, a precipitate already forms. The mixture is stirred for 2 hours, then filtered and air dried to give 6.5 g of amide. 1.4 g (10 mmol) of the abovementioned compound and 1.5 g ketobutyric acid (15 mmol) are refluxed in 25 ml toluene for 15 hours to remove water. The reaction mixture is cooled and partially evaporated with a stream of _N2 . Crystallization of the product occurs after standing for 3 hours. The crystals are collected and washed three times with toluene and air dried. 1.13 g (50%) of product, melting point 160-162°, is isolated. NMR is consistent with the assigned structure and shows 5% E isomer. TLC (4:1
Toluene-AcOH) shows a single spot. Analysis (C ₁₂ H ₂₁ NO ₃ ) Calculated value Actual value C 63.40 63.63 H 9.30 9.43 N 6.16 5.88 Example 10 Z-2-(2,2-dimethylcyclopropanecarboxamide)-2-butenoic acid, 1.53 g (15 mmol) ) 2-ketobutyric acid and 1.13g
(10 mmol) of 2,2-dimethylcyclopropanecarboxamide and 20 ml of toluene are stirred under reflux for 10 hours. After cooling, the crystalline solid is filtered, washed with toluene (1 x 10 ml) and dried to give 1.06 g of product. Melting point 140-141℃. TLC (4:
1 toluene-AcOH) shows essentially one spot and the NMR spectrum is consistent with the desired structure. Recrystallization from EtOAc and drying yielded 0.533 g of product, mp 142-143.5°, single by TLC. Analysis C ₁₀ H ₁₅ NO ₃ Calculated value Actual value C 60.90 60.92 H 7.67 7.71 N 7.10 7.38 Example 11 Z-2-(2,2-dimethylcyclopropanecarboxamide)-2-hexenedionic acid, 1.0 g of 2,2 -dimethylcyclopropanecarboxamide, 2.4 g of 2-ketoadipic acid and 25 ml
A mixture of methyl isovalerate is heated to reflux for 4 hours and dehydrated in a modified Dean-Stark trap containing molecular sieves (4A). After standing overnight at room temperature, the crystalline precipitate was filtered, washed with ether, and recrystallized from ethyl acetate to yield 0.23 g.
A product with a melting point of 163-165° is obtained. The NMR spectrum is consistent with the desired structure. Analysis (C ₁₂ H ₁₇ NO ₅ ) Calculated value Actual value C 56.46 56.20 H 6.71 6.83 N 5.49 5.32 Example 12 Z-2-(2,2-diethylcyclopropanecarboxamide)-2-butenoic acid 2.3 g of 2-keto A mixture of butyric acid, 2.0 g of 2,2-diethylcyclopropanecarboxamide and 25 ml of toluene is heated to reflux for 16 hours and the water removed in a modified Dean-Stark trap containing molecular sieves (4A). The product does not precipitate on cooling. Add ether (25ml) to saturate this mixture
Extract 3 times with _NaHCO3 . Combine the extracts and acidify with concentrated HCl. A gummy precipitate crystallizes on treatment with water. Recrystallization from ethyl acetate gives 0.31 g of product, mp 129-130°. The NMR spectrum is consistent with the desired structure. Analysis (C ₁₂ H ₁₉ NO ₃ ) Calculated value Actual value C 63.98 64.01 H 8.50 8.62 N 6.22 6.21 Example 13 2-(2,2-dimethylcyclopropanecarboxamide)-2-hexenoic acid Step A DL-norleucine t-butyl Esters General Procedures, R.Roesske, J.Org.Chem, Volume 28
Page 1251 (1963) in a 500 ml pressure bottle cooled in an ice bath.
9.82 g (75 mmol) in dioxane in 80 ml
Add 8 ml of concentrated H ₂ SO ₄ to DL-Norleucine.
Add. Add 80 ml of liquid isobutylene while cooling the resulting mixture in a dry ice bath.
The mixture is warmed to room temperature and shaken under autogenous pressure for ~23 h. After evacuating most of the isobutylene, the slightly hazy solution is cooled in ice and then mixed with 400 ml of
Add to a cold mixture of 1N NaOH and 500 ml _Et2O . After shaking in a separatory funnel, the two layers are separated and the aqueous layer is washed with an additional 100 ml of Et ₂ O. 150ml of _Et2O solution
Shake with 0.5NHCl. The acidic aqueous solution was treated with 2.5N NaOH until strongly basic, then 250ml of _Et2O
Toful. The Et ₂ O solution was dried (MgSO ₄ ) and filtered.
Concentrate on a rotary evaporator. After long periods of pumping under high vacuum over a steam bath, a clear, clear oil remains at the end. 9.04 g (65%), NMR shows only traces of dioxane remaining.
TLC (9:1 CHCl ₃ -MeOH) shows a single spot. Step B N-(2,2-dimethylcyclopropanecarbonyl)-DL-norleucine t-butyl ester Attached to drying tube and stirred in dry ice bath
8.98 g (48 mmol) DL− in 100 ml CH ₂ Cl ₂
norleucine t-butyl ester and 5.05 g (50 mmol) of triethylamine solution in 50 ml of
6.39 g (48 mmol) chloride 2 in CH ₂ Cl ₂ ,
2-dimethylcyclopropanecarbonyl (M.
Elliot and NRJames, British Patent No. 1260847 1972
2)) Add the solution dropwise (over 75 minutes). Precipitation of Et ₃ NHCl occurs during the addition, especially towards the end. As the ice gradually melts, allow the temperature of the mixture to rise to room temperature. After 16 hours, this mixture
200ml of 0.5NHCl and sifter. CH ₂ CL ₂ classification further 200
ml of 0.5NHCl, then 2 x 200ml of 0.5NNaOH,
Finally, wash with 200ml of water. CH ₂ Cl ₂ divisions
Dry with _MgSO4 , treat with activated charcoal, and pass through Celite. Concentrate the liquid using a rotary evaporator (high vacuum at the end). Bright orange residual oil = 11.93g (88%). TLC (2:1 hexane-EtoAc) shows a single spot. NMR and
IR is consistent with the assigned structure. After a few days, the unused portion of the substance crystallized. melting point
52−>65°. Step C t-Butyl 2-(2,2-dimethylcyclopropanecarboxamide)-2-methoxyhexanoate H. Poisel and V. Schmidt, Chem. Ber. p. 108,
Based on page 2547 (1975). Stir 35 ml at room temperature in the dark under a stream of _N2 .
6.37 g (22.5 mmol) of N-( _2,2
-dimethylcyclopropanecarbonyl)-DL-norleucine t-butyl ester solution in 2.69ml
(2.45 g, 22.5 mmol) of t-butylpipochlorite is added. After 15 minutes, 0.52 in 35ml MeOH
A sodium methoxide solution prepared by dissolving 22.6 mmol of sodium is added. Continue stirring at room temperature in the dark under a stream of _N2 .
After 16.5 hours, the precipitated NaCl is removed. liquid
Dilute with Et ₂ O, wash with 3 x 50 ml of 0.5NHCl, 50 ml of saturated Na ₂ CO ₃ and 2 x 50 ml of water. Concentrate the Et ₂ O layer on a rotary evaporator. The light golden residual oil (6.45 g) was subjected to preparative high-pressure liquid chromatography to produce two compounds, 273 mg and 496 mg of t-butyl 2-(2,2-dimethylcyclopropanecarboxamide)-2-methoxyhexanoate. diastereomers (melting points 114-118° and 124-125.5°, respectively) and 1.97 g of t-butyl 2-
A single isomer of (2,2-dimethylcyclopropanecarboxamide)-2-hexenoate (apparently Z, colorless oil) is obtained. Step D 2-(2,2-dimethylcyclopropanecarboxamide)-2-hexenoic acid 0.84 g in 10 ml _Et2O saturated with anhydrous HCl
A solution of (3.0 mmol) in t-butyl 2-(2,2-dimethylcyclopropanecarboxamide)-2-hexenoate is left at room temperature under a drying tube. After 17 hours, the solution is evaporated and the remaining gum is dissolved in 10 ml of saturated _NaHCO3 . Add this solution further
Wash with 15 ml of 0.5NHCl and then dry (MgSO ₄ )
Filter and concentrate to obtain a viscous oil. This oil is crystallized from toluene. Yield of white crystals = 0.32g
(47%), mp 119-122°. TLC (4:1 toluene-AcOH) showed a single spot. NMR
indicates substantially pure Z-isomer. (Note: methanol adduct, t-butyl 2-
(2,2-dimethylcyclopropanecarboxamide)-2-methoxyhexenoate under the same conditions.
Treatment with anhydrous HCl in _Et2O gives the same product). Example 14 (+)-Z-2-(2,2-dimethylcyclopropanecarbonylamino)-2-octenoic acid sodium salt Reagent, (+)-2,2-dimethylcyclopropanecarboxamide, 7.0 g; 2-keto 14.7 g of octanonic acid ethyl ester; 50 mg of P-toluenesulfonic acid; 100 ml of toluene are placed in a 250 ml three-neck flask equipped with a Dean-Stark trap containing several molecular sieve pellets. This mixture is refluxed vigorously for 27 hours. The resulting bright yellow solution is cooled and concentrated under vacuum in a 45° C. water bath with the addition of water to remove toluene. The cam-like residue is suspended in 230 ml of 2N NaOH and stirred at 30°C for 3 hours, then the temperature is increased to 35°C for an additional 21/2 hours until a clear liquid is obtained. The solution is then cooled, 85 ml of methylene chloride is added and the pH is adjusted to 8.5 by adding 4N HCl while stirring.
Separate the organic layer and discard. When the aqueous layer (366 ml) was analyzed by liquid chromatography, it was 37.2 mg/ml; 87
% of the Z isomer. Then add another 85 ml of CH ₂ Cl ₂ and adjust the pH to 4.5 while stirring. The organic layer is separated and the aqueous layer is adjusted to PH4.5 again and extracted with 50 ml of CH ₂ Cl ₂ . Combine the organic layers, dry over Na ₂ SO ₄ , filter, and concentrate to a cam. Mix this residue with 150ml of isopropanol.
Dissolve in 15 ml of water and adjust the pH to 8.2 with 2N NaOH. The resulting solution is concentrated to an oily residue and flash concentrated with isopropanol until the residue turns into a crystalline solid (this means that most of the water has been removed). This solid substance
Crystallize from 120 ml of isopropanol (chilled in ice for 1 hour), filter and wash with 50 ml of cold isopropanol and then copious amounts of acetone. This is 60
Drying for 2 hours at °C/0.1 mm gives 10.74 g (63.2% Kl) of a crystalline material giving essentially a single peak in liquid chromatography, melting point 241-243 °C. The starting material, (+)-2,2-dimethylcyclopropanecarboxamide, was obtained by optically resolving the D,L acid.
Most conveniently, the optically resolved amide is prepared by reaction with oxalyl chloride followed by ammonia. One method of preparing the starting material is as follows: 23.1 g of D,L-2,2-dimethylcyclopropanecarboxylic acid is suspended in 33 ml of water and 50%
Adjust the pH to 8.0 using approximately 10 ml of NaOH. 30
Pre-add about 8 ml of concentrated HCl in 1 ml of water to bring the pH to 7.1.
A solution of 38.4 g of quinine (which is actually a quinine hydrochloride solution) in a mixture of 60 ml of methanol and 30 ml of water that has been prepared is first added to the solution. Add these solutions all at once while stirring. When the resulting gum is heated, two transparent layers are obtained, and when it is vigorously stirred again while cooling, a crystalline material is obtained. The product is left at room temperature for 2 days. Then strain 2 x 10 ml water, 2 x 10 ml
Wash with 50% ethanol and air dry with suction. Yield of crude quinine salt: 44.8g (48.7% yield),
Monohydrate. Melting point 113-116℃, [α] ^20D _is -94.3℃,
(C=1.0 in _CHCl3 ). Recrystallization of this material from acetone yields 24.35 g, mp 127-130°C. This purified quinine salt is converted to acid by reacting with alkaline water and chloroform and then with acid, yield (96%), 3.9
g, [α] ²⁰ _D +146.0° This acid is converted to an amide as follows. Add 30.5 g of (+) acid through an addition funnel to 54 ml of chilled (10° C.) oxalyl chloride containing 1 drop of dimethylformamide over 5-10 minutes. This is stirred at room temperature overnight. This will be a clear liquid, so add it to 100ml of methylene chloride to dilute it. Remove excess oxalyl chloride by concentration,
The mixture is flash concentrated twice with methylene chloride. The resulting solution is diluted with an equal volume of methylene chloride, added successively through an addition funnel to about 100 ml of anhydrous liquid ammonia, and diluted with 100 ml of methylene chloride. A dry ice-acetone cooling bath is used during the addition. Once all is added, remove the cooling bath and stir the mixture at room temperature for about 1/2 hour. Filter the mixture to remove the liquid ammonium precipitate;
Concentrate and dry. The weight of the crude product is 26.6g (88%)
It is. This was dissolved again in excess hot ethyl acetate and filtered through a preheated glass filter.
Separate from trace amounts of NH ₄ Cl. Excess ethyl acetate is distilled off under atmospheric pressure. When the liquid volume is reduced to half,
Add 130 ml of heptane and continue to distill off the ethyl acetate until the boiling point starts to rise (to around 80°C, most of the product has already precipitated out). Heating is discontinued and the mixture is slowly cooled to about 30°C and then cooled in an ice bath at 0-5°C for about 1/2 hour. The product was recovered as clean silvery white crystalline flakes and
Wash with xethyl acetate/hexane mixture, 1/1.5, and air dry to constant weight. Weight is 23.3g
(total yield 77.1%, recovery rate from crude product 87.6%), and the melting point is 135-138°C (varies depending on the heating rate). The optical rotation was determined as [α] ²⁰ _D = +100.9° by dissolving 0.0543 g in 10 ml of chloroform. Example 15 Z-2-(2,2-dichlorocyclopropanecarboxamide)-2-butenoic acid Step A: 2,2-dichlorocyclopropanecarboxamide 7.1 g of 2,2-dichlorocyclopropanecarbonyl chloride (U.S. Pat. No. 3,301,896) Issue, January 1967
31st issue) into 75 ml of concentrated ammonium hydroxide while stirring vigorously. Keep the temperature of the reaction mixture below 10°C using an ice bath. Mix this mixture in an ice bath for 30 min.
Stir for 1 minute and then 1 hour at room temperature. Evaporate the aqueous ammonia under reduced pressure. (Bath temperature 50℃). The solid residue is extracted with hot ethyl acetate (3 x 30 ml).
Concentrate the extract to 40 ml and add 20 ml hexane. After cooling in ice, the solid was filtered, washed with ethyl acetate-hexane (1:1) and dried.
2.7 g of 2,2-dichlorocyclopropanecarboxamide, melting point 144-146°, are obtained. The NMR spectrum is consistent with the desired structure. Analysis (C ₁₂ H ₅ Cl ₂ NO) Theoretical value Actual value C 31.20 31.26 H 3.27 3.31 N 9.10 9.11 Cl 46.04 45.79 An additional 1.3 g of amide, melting point 143-145°, can be recovered from the mother liquor. Step B: Z-2-(2,2-dichloropropanecarboxamide)-2-butenoic acid 1.53 g (15 mmol) of 2 in 10 ml of toluene
- a mixture of ketobutyric acid and 1.54 g (10 mmol) of 2,2-dichlorocyclopropanecarboxamide
Heat to reflux for an hour and remove water with a modified Dean Stark trap containing molecular sieves (4A).
A further 0.7 g of 2-ketobutyric acid is added and the reaction mixture is heated to reflux for a further 12 hours. Cool this mixture 20
Dilute with ml of toluene and saturated sodium bicarbonate water (3 x 10ml)
Extract with Combine the extracts, wash with ether, and acidify to PH3 (PH meter) with concentrated hydrochloric acid. A gum precipitates and quickly solidifies. This was filtered, washed with water, dried and reconstituted from nitromethane to yield 423 mg of Z-2-(2,2-dichlorocyclopropanecarboxamide)-2-butenoic acid, mp.
188-189.5℃ is obtained. The NMR spectrum is consistent with the desired structure. Analysis (C ₈ H ₉ Cl ₂ NO ₃ ) Calculated value Actual value C 40.36 40.38 H 3.81 3.80 N 5.88 5.91 Cl 29.78 29.53 Example 16 Z-2-(2,2-dichlorocyclopropanecarboxamide)-2-octenoic acid 1.19 g (7.5 mmol) of 2-ketooctanoic acid, 0.77 g (5.0 mmol) of 2,2-dichlorocyclopropanecarboxamide and 5 ml of toluene are reacted according to the same procedure as in the previous example. The crude product (537 mg) was converted to methyl ester (BF ₃ /CH ₃ OH) and subjected to preparative TLC (silica gel G,
4:1 hexane-EtOAC) to give pure 2
-Saponification of methyl ester (0.3MLiOH/
_CH3OH ) 88 mg of Z-2-(2,2-dichlorocyclopropanecarboxamide)-2-octenoic acid are obtained as a partially crystalline gum.
NMR spectrum (DMSO- _d6 ): 9.68δ(s,
1H, NH), 6.50δ (t, 1H, [Formula] 2.83δ (t, 1H, [Formula]), 1.97δ (d, 2H, [Formula]) Example 17 Z-8-Bromo-2-(2 ,2-dimethylcyclopropanecarboxamide)-2-octenoic acid into a suspension of 14.4 g (0.3 mol) of a 50% NaH dispersion in 360 ml of toluene under an _N2 atmosphere for 45 min, cooled in an ice bath. 146 in 120ml DMF over
g (0.6 mol) of 1,6-dibromohexane and
A solution of 57.6 g (0.3 mol) of ethyl 1,3-dithiane-2-carboxylate is added. The cooling bath is removed and the mixture is stirred at room temperature for 2 hours. The reaction mixture was washed with water (3 x 210 ml), dried over _MgSO4 and evaporated under reduced pressure to give the desired anhydrous dithiane, 1,
179.5 containing 6-dibromohexane and mineral oil
g of yellow oil is obtained. This crude material is used in the next reaction without purification. 426g in 800ml acetonitrile and 200ml water
A solution of crude dithiane in 100 ml of acetonitrile is added to a suspension of (2.4 mol) of N-bromosuccinamide over a period of 45 minutes. The temperature of the reaction mixture is kept below 25°C with an ice bath. After stirring for 10 min at 20°C, the dark red reaction mixture was diluted with 2 hexane-
Pour into CH ₂ Cl ₂ (1:1). Then add 400 ml of saturated Na ₂ Cl ₃ solution in small portions (CO ₂ is generated vigorously)
Add. After foaming is complete, shake with a funnel to separate the aqueous layer. The organic layer is extracted with saturated Na ₂ CO ₃ solution (400 ml) and water (500 ml) and dried over MgSO ₄ . Removal of the solvent under reduced pressure yields 133.8 g of crude bromoketoester containing 1,6-dibromohexane and mineral oil. Use this crude material in the next reaction without purification. A mixture of 133.8 g of crude bromoketo ester, 133 ml of hydrobromic acid and 267 ml of acetic acid is heated at 90° C. (internal temperature) for 75 minutes. The dark solution is evaporated under reduced pressure until most of the acetic acid is gone. residue
Dissolve in 500 ml of ether and wash with water (2 x 100
ml) and extracted with saturated _NaHCO3 (3 x 200 ml). The combined _NaHCO3 extracts are extracted with ether (2 x 100 ml) and acidified with concentrated hydrochloric acid. The precipitated oil is extracted with ether (3 x 200 ml). The ether extract is washed with water (1 x 100 ml) and saturated brine (1 x 100 ml) and dried over _MgSO4 . Removal of the ether under reduced pressure yields 46.2 g of pure bromomoketo acid. Single by TLC (silica gel, 4:1 toluene-acetic acid). The NMR spectrum is consistent with the desired product. 46.1 g (0.194 mol) Bromo Keto Acid 17.6 g
(0.156 mol) of 2,2-dimethylcyclopropanecarboxamide and 450 ml of toluene are heated to reflux for 13 hours, while water is collected in a small Dean-Stark trap. After cooling, the clear reaction mixture is extracted with saturated NaHCO ₃ solution (4×100 ml). The combined extracts are washed with ether (2 x 100 ml) and then concentrated HCl is added to adjust the pH to 3.5 (PH meter). An oil precipitates, which quickly crystallizes. Filter this solid material, wash thoroughly with water, and dry. When reconstituted from acetonitrile, 22.5 g of Z-
8-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid, melting point 151-153°C, is obtained. Homogeneous by TLC (4:1 toluene-acetic acid). The NMR spectrum is consistent with the desired structure. Analysis (C ₁₄ H ₂₂ BrNO ₃ ) Calculated value Actual value C 50.61 50.66 H 6.67 6.96 N 4.22 4.45 Br 24.05 23.95 The following ω-bromo compounds are also prepared by the same method. Z-6-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-hexenoic acid; Z-7-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-heptenoic acid; Z-9-bromo- 2-(2,2-dimethylcyclopropanecarboxamide)-2-nonenoic acid; Z-10-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-decenoic acid; Z-8-bromo-2-(2 , 2-dimethylcyclopropanecarboxamide)-octenoic acid. Example 18 Z-8-dimethylamino-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid 664 mg (2 mmol) of Z-8-bromo-2- in 10 ml of 40% dimethylamine solution The (2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid solution is left at room temperature for 4 hours. Pour this solution onto a 3.5 x 20 cm column of Dowex 50W-X8 (100-200 mesh, H ⁺ ) ion exchange resin and wash with water (~200
ml) to elute this column. Then column 300
Elute with ml of 2N ammonium hydroxide. Evaporation of the effluent under reduced pressure gives 600 mg of colorless glass. This material is dissolved in 3 ml of ethanol, filtered and added dropwise to 200 ml of rapidly stirring acetone. A gummy solid precipitates, but
This will crystallize if left for 2 days. When this solid substance is filtered, washed with acetone, and dried, 445 mg of Z
-8-Dimethylamino-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid is obtained as colorless, hygroscopic crystals with a melting point of 101 DEG -112 DEG C. TLC (silica gel, BuOH,
HOAc, H ₂ O4: 1:1). The NMR spectrum is consistent with the desired structure. Analysis _{( C16H28N2O8.H2O} ) Calculated Values Observed Values C 61.12 _61.03 H 9.62 9.28 N 8.91 8.67 The following 8-amino derivatives are prepared by substantially _{the same} method. "DCC" means 2-(2,2-dimethylcyclopropanecarboxamide). Z-10-dimethylamino-DCC-2-decenoic acid; Z-8-amino-DCC-2-octenoic acid; Z-8-dimethylamino-DCC-2-octenoic acid; Z-7-dimethylamino-DCC- 2-heptenoic acid; Z-DCC-7-(N-methylpiperazinyl)-
2-heptenoic acid; Z-DCC-8-pyrrolidino-2-octenoic acid; Z-DCC-8-(N-methylpiperazinyl)-
2-octenoic acid; Z-8-allylamino-DCC-2-octenoic acid; Z-DCC-8-piperidino-2-octenoic acid; Z-DCC-8-propargylamino-2-octenoic acid; Z-8-N -[1-deoxy(1-methylamino)-D-glucityl]-DCC-2-octenoic acid; Z-8-(1-adamantylamino)-DCC-
2-octenoic acid; Z-8-diallylamino-DCC-2-octenoic acid; Z-8-DCC-8-(2-hydroxyethylamino)-2-octenoic acid; Z-8-[(carboxymethyl)methyl amino〕
-2-(2,2-DCC)-2-octenoic acid; Z-2-(2,2-DCC)-8-diethylamino-2-octenoic acid; Z-2-(2,2-DCC)-8 -[Tris-(hydroxymethyl)methylamino]-2-octenoic acid; Z-2-(2,2-DCC)-10-(N-methylpiperazinyl)-2-octenoic acid; Z-2-( 2,2-DCC)-8-[1-(phosphono)ethylamino]-2-octenoic acid Z-8-[(carboxymethyl)methylamino]
-2-(2,2-dimethyl-cyclopropanecarboxamide)-2-octenoic acid Z-B-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid 3.32 g, CH ₃ NHCH ₂ CO ₂ H1.0g, Na ₂ CO ₃ 3.5g
and 30 ml of water were heated at 80 °C for 1.5 h in _N2 .
After purification, 1.0 g of product was obtained. Analysis (C ₁₇ H ₂₈ N ₂ O ₅・2H ₂ O) Calculated value Actual value C 54.24 54.40 H 8.57 8.34 N 7.44 7.16 Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-[1-(phosphono ) ethylamino]-2-octenoic acid The title compound is the same bromo intermediate (335.1mg)
was prepared by reacting with 138.2 mg of 1-aminoethanephosphoric acid and 435 mg of Na ₂ CO ₃ in 5 ml of water and then following essentially the same procedure. Ki=0.16 Z-2 [(1S)-2,2-dimethylcyclopropanecarboxamide]-8-[(1RS)-1-(phosphono)-ethylamino]-2-octenoic acid This compound is (+)-Z -8-Bromo-2-
(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid was prepared by the method described above but obtained as a crystalline solid. mp143−145°, [α] ²⁰ _D = +12.54° (H ₂ O, 1). Example 19 Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-methylthio-2-octenoic acid in 5 ml of methanol cooling in an ice bath.
A solution of 162 mg (3 mmol) of sodium methoxide is injected with a stream of CH ₃ SH for 10 minutes. The solution is brought to room temperature and 332 mg (1 mmol) of Z-8-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid are added. Heat the solution to reflux for 30 min under _N2 atmosphere. Evaporate most of the methanol under reduced pressure and reduce the residue to 10
Dissolve in ml of water and acidify with 2.5NHCl. Extract the precipitated oil with ether (3X). Wash the ether extract with water, saturated brine, and dry over _MgSO4 . Removal of the ether under reduced pressure yields a colorless oil;
If left alone, it will crystallize. Reconsolidation from ether-hexane gives 178 mg of Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-methylthio-2-octenoic acid, mp 82-84°C. Homogeneous by TLC (toluene-acetic acid, 4:1). The NMR spectrum is consistent with the desired structure. Analysis (C ₁₅ H ₂₅ NO ₃ S) Calculated value Actual value C 60.18 60.36 H 8.42 8.68 N 4.68 4.59 S 10.69 10.87 The following compounds are prepared in a similar manner.
"DCC" means 2-(2,2-dimethylcyclopropanecarboxamide). Z-DCC-8-ethoxythiocarbonylthio-2-octenoic acid; Z-DCC-8-(1-methyl-5-tetrazolylthio)-2-octenoic acid; Z-DCC-7-{[(methoxycarbonyl)methyl ]thio}-2-heptenoic acid; Z-8-acetylthio-DCC-2-octenoic acid; Z-7-[(2-amino-2-oxoethyl)thio]-DCC-2-heptenoic acid 6-(L- 2
-amino-2-carboxyethylthio)-2-
(2,2-DCC)-2-hexenoic acid; Z-8-(carbomethoxymethylthio)-2-
(2,2-DCC)-2-octenoic acid; Z-6-(carbomethoxymethylthio)-2-
(2,2-DCC)-2-hexenoic acid; Z-2-(2,2-DCC)-6-(phosphonomethylthio)-2-hexenoic acid; Compound 7-(L-2-amino-2- Carboxyethylthio)-2-(2,2-dimethylcyclopropanecarboxamide)-2-heptenoic acid is prepared in a manner similar to the previous example. However, Z-7
-bromo-2-(2,2-dimethylcyclopropanecarboxamide)-2-heptenoic acid
Dissolved in 2.02ml of NaOH solution (2.ON) (185mg,
1.05 mmol) and deoxygenated by bubbling a stream of nitrogen gas through the solution for 1 minute.
Next, cysteine/HCl (185 mg, 1.05 mmol)
are added all at once and the reaction is stirred at room temperature under _N2 atmosphere for 3 hours. The mixture was then added with 300ml of _H2O
Elute with 200 ml of _2NNH3 solution. Ammonia is distilled off under reduced pressure to obtain 284 mg of yellowish glass. This product is dissolved in 4 ml of ethanol and the insoluble material is filtered off. Add the solution dropwise to rapidly stirred diethyl ether (150 ml). Filter the precipitated solid, wash with ether and dry to give the product.
Get 171mg. The product was purified by thin layer chromatography (TLC) (nBUOH, HOAC, H ₂ O; 4:1:
1) gives 1 spot (ninhydrin positive),
rf. about 6; NMR is good. Analysis (C ₁₆ H ₂₆ N ₂ O ₅ S) Calculated Value Found C 53.61 52.55 H 7.31 7.40 N 7.81 7.89 S 8.94 9.63 The following compounds were prepared using the same method. 7
-(D-amino-2-carboxyethylthio)-2
-(2,2-dimethylcyclopropanecarboxamide)-2-heptenoic acid was obtained as a crystalline solid. Melting point 183-185° (decomposed). Homogeneous by thin layer chromatography [silica gel;
nBUOH, HOAC, H ₂ O (4:1:1); ninhydrin positive], the NMR spectrum was consistent with the desired structure. Ki=0.19μM Analysis (C ₁₆ H ₂₆ N ₂ O ₅ S・25H ₂ O) Calculated value Actual value C 52.95 53.04 H 7.36 7.25 N 7.72 7.43 S 8.83 9.02 6-(L-2-amino-2-carboxyethylthio )-2-(2,2-dimethylcyclopropanecarboxamide)-2-hexenoic acid was obtained as a crystalline solid. Melting point 120-132° (decomposed). Thin layer chromatography [silica gel; nBUOH,
Homogeneity due to HOAC, H ₂ O (4:1:1); ninhydrin positive]. The NMR spectrum was consistent with the desired structure. Ki=0.27μM Analysis (C ₁₅ H ₂₄ N ₂ O ₅ S・5H ₂ O) Calculated value Actual value C 50.95 50.86 H 7.13 7.11 N 7.93 7.76 S 9.07 9.25 8-(L-2-amino-2-carboxyethylthio )-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid was obtained as a crystalline solid. Melting point 174-177° (decomposed). Thin layer chromatography [silica gel; nBUOH,
Homogeneity due to HOAC, H ₂ O (4:1:1); ninhydrin positive]. The NMR spectrum was consistent with the desired structure. Ki = 0.23μM. Z-7-(L-amino-2-carboxyethylthio)-2-(2,2-dimethylcyclopropanecarboxamide)-2-heptenoic acid sodium A Grignard production of ethyl-7-chloro-2-oxoheptanoate Equimolar amounts of 1-bromo-5-chloropentane and magnesium (8 moles each) are reacted in tetrahydrofuran (960 ml) at 25°C. Pour a solution of Mg in THF into a flask. Then add bromochloropentane over 1 hour and then leave for 2 hours. After the reaction was determined to be complete, the reaction solution was cooled to -15°C and the temperature was lowered to -15°C.
1856ml of tetrahydrofuran while maintaining at 10℃
Add to 16 moles of diethyl oxalate in the solution.
Quench by adding 3NHCl and keep the temperature below 25 °C. After stripping the solvent, the calculated yield was ethyl-1-chloro-6-oxoheptanoate.
It is 48.8%. B Condensation and Hydrolysis S-2,2-dimethylcyclopropylcarboxamide (1017g) Ethyl-7-chloro-2-
2143.6 g of ketoheptanoate, 9 g of toluene and 12 g of p-toluenesulfonic acid were placed in a 22 liter flask and heated to reflux with stirring.
After 23 hours, liquid chromatography showed the expected product ratio. Toluene 4 was then removed under light vacuum. pour water into the pot
Neutralized to pH 7 with 2N NaOH and distilled under reduced pressure to a final pot volume of approximately 5. This was hydrolyzed by adding 1760 g of 50% aqueous NaOH (4 ml of water) and stirred overnight. Add 4 methylene chloride to the flask and adjust the pH to 8.8 using HCl.
It was adjusted to Unreacted amide crystallized. The organic layer was separated from water and then evaporated. gummy residue
Dissolve 720 g of 50% NaOH in 8 g of water, add 1818 g of L-cysteine HCl.H ₂ O to this solution and add 2 g of ice.
Kg, 2484 g of 50% NaOH and 1 portion of water were added. After standing overnight at room temperature, the pH of the solution was adjusted to 3.0 with concentrated hydrochloric acid.
The resulting gummy suspension is heated to 95°C to give a clear solution. After 30 minutes no E isomer was detected by _IC . After work-up and purification the overall yield was 50%. This material was recrystallized from acetonitrile. For the recrystallized material, 1500g is mixed with water 6 and
Dissolve in 910ml of 3.88NNaOH and then neutralize to PH
7 and pear, lyophilized to yield 1569 g of the title compound
(98.6%). Analysis Calculated value Actual value C 50.52 50.71 H 6.62 6.78 N 7.36 7.49 S 8.43 8.52 Na 6.04 5.92 Z-8-[(2-amino-2-oxoethyl)thio]-2-(dimethylcyclopropanecarboxamide)-2-octenoic acid The title compound was also prepared in a similar manner as described in Example 19 with 3.3 g of bromo intermediate 50 g of methanol
Prepared using 1.3 g of H ₂ NC (=0) CH ₂ SH in ml, yielding 1.6 g of product. Melting point 127-128℃. Example 20 Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-trismethylammonium hydroxide-2-octenoic acid inner salt 996 mg in 15 ml of 28% triethylamine aqueous solution
(3 mmol) of Z-8-bromo-2-(2,2-
dimethylcyclopropanecarboxamide)-2-
The octenoic acid solution is left at room temperature for 3 hours. Convert the reaction mixture to IRA-410 (50-100 mesh, OH ^- )
Pour onto a 2 x 25 cm column of ion exchange resin and elute with water until the effluent is no longer basic.
Evaporation of the effluent under reduced pressure gives 800 mg of a colorless glass. Dissolve this material in 20 ml of ethanol, filter, and dilute with 600 ml of acetone. After standing overnight at room temperature, the precipitated crystalline solid was filtered, washed with acetone, and dried to yield 720 mg.
Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-trimethylammonium hydroxide-2-octenoic acid inner salt is obtained as hygroscopic crystals, melting point 220-222°C. Homogeneous by TLC (silica gel, BuOH, HOAc, H ₂ O, 4:1:1). The NMR spectrum is consistent with the desired structure. Analysis (C ₁₇ H ₃₀ N ₂ O ₃ ) Calculated Value Found C 65.77 65.78 H 9.74 9.98 N 9.02 8.92 Other quaternary derivatives were prepared using substantially the same method. These are Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-trimethylammonium hydroxide-2-octenoic acid inner salt; Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-pyridinium hydroxy Do-2-octenoic acid inner salt; Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-(2-hydroxydimethylammonium hydroxide)-2-octenoic acid inner salt; Z-2-(2, Z-8-(benzylmethylammonium hydroxide)-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoin Acid inner salt; Z-10-(benzyldimethylammonium hydroxide)-2-(2,2-dimethylcyclopropanecarboxamide)-2-decenoic acid inner salt; Z-2-(2,2-dimethylcyclopropanecarboxamide) -9-trimethylammonium
Hydroxide-2-noneinic acid inner salt; Z-8-(2-dimethylaminoethyldimethylammonium hydroxide)-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid inner salt; Z-2- (2,2-dichlorocyclopropanecarboxamide)-8-trimethylammonium
Hydroxide-2-octenoic acid inner salt; Example 21 Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-formamidino-2-octenoic acid 350 mg of Z-8-amino-2-(2, Dissolve 2-dimethylcyclopropanecarboxamide)-2-octenoic acid in 10ml of water and add 2.5NNaOH to pH 8.5.
Adjust to A total of 9.47 mg of benzylformimidate hydrochloride is added in portions over 20 minutes at room temperature while maintaining the pH at 8-9 by adding 2.5 N NaOH. After stirring at room temperature for 30 min, the cloudy reaction mixture was extracted with ether (3x) and the G50W-X4
Apply to a 2 x 25 cm column of (Na ⁺ , 200-400 mesh) resin. After elution with water, the fractions containing the product are collected and evaporated under reduced pressure. Dissolve this substance in water G1 x 8 (HCO ₃ ^- , 200-400 mesh)
Apply to a 2 x 25 cm column of resin. After elution with water,
The fractions containing pure product are collected and evaporated under reduced pressure. The residue is dissolved in a few ml of warm ethanol, filtered and added dropwise to 200 ml of ether with rapid stirring. 243mg when filtered and washed with ether
Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-formamidino-2-octenoic acid is obtained as an amorphous solid.
TLC (n-BuOH, HOAc, _H2O , 4:1:1)
It is uniform. The NMR spectrum is consistent with the desired structure. Analysis (C ₁₅ H ₂₅ N ₃ O ₃ ·1/3H ₂ O) Calculated value Actual value C 59.69 60.04 H 8.59 8.64 N 13.92 13.57 The following amidino compounds were prepared in a similar manner. Z-8-acetamidino-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid; Z-8-benzylamidino-2-(2,2-dimethylcyclopropanecarboxamide-2-octenoic acid; Z- 2-(2,2-dimethylcyclopropanecarboxamide)-10-formamidino-2-decenoic acid; Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-(2-imidazoyl-2-yl-amino )-2-octenoic acid Example 22 Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-guanidino-2-octenoic acid 2 mmol guanidine (432 mg
of guanidine sulfate and 630 mg of barium hydroxide.
332 mg (1 mmol) of 8-bromo-2-(2,2-dimethylcyclopropanecarboxamido)octenoic acid are added to the solution (prepared from 8-hydrate) and the solution is heated at 70° C. for 1 hour under a nitrogen atmosphere.
Dowex 50W−X8 (H ⁺ , 100−
Pour onto a 2 x 25 cm column of 200 mesh. After elution with water, the fractions containing the product are collected and evaporated under reduced pressure. The residue is dissolved in a few ml of warm ethanol and added dropwise to 100 ml of ether with rapid stirring. After filtering and washing with ether, 107 mg of Z-
2-(2,2-dimethylcyclopropanecarboxamide)-8-guanidino-2-octenoic acid is obtained as an amorphous electrostatic powder. TLC
It is uniform (n-BuOH, HOAc, H ₂ O; 4:1:1). NMR (D ₂ O, NaOD): 6.48δ (t,
1H, [Formula]); 3.10δ (m, 2H, [Formula]), 2.10δ (m, 2H, [Formula]) 1.17δ (s, 3H, [Formula]) The following guanidino compounds were prepared by the same method. did. Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-(N,N-dimethylguanidino)-2-octenoic acid. Example 23 Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-methoxy-2-octenoic acid 332 mg (1 mmol) of 8 in a solution of 2.43 mmol sodium methoxide in 5 ml methanol.
-Bromo-2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid is added. This solution is heated to reflux under a nitrogen atmosphere for 1 hour. The reaction mixture is evaporated under reduced pressure and the residue is dissolved in water and acidified with 2.5N hydrochloric acid. The precipitated oil is extracted with ether (3x). The ether extract was washed with water, saturated brine and dried over _MgSO4 . Removal of the ether under reduced pressure gives a colorless oil which crystallizes on standing. When this was reconstituted from ether-hexane, 140 mg of Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-methoxy-
2-octenoic acid, melting point 71-72°C, is obtained.
Homogeneous by TLC (toluene-HOAc, 4:1). The NMR spectrum is consistent with the desired structure. Analysis (C ₁₅ H ₂₅ NO ₄ ) Calculated value Actual value C 63.58 63.54 H 8.89 9.12 N 4.94 5.16 The following compounds were prepared using a similar method. Z-8-cyano-2-(2,2-dimethylcyclopropanecarboxamide-2-octenoic acid; Z-7-cyano-2-(2,2-dimethylcyclopropanecarboxamide)-2-heptenoic acid; Z-9 -Cyano-2-(2,2-dimethylcyclopropanecarboxamide)-nonenoic acid; Z-2-(2,2-dimethylcyclopropanecarboxamide)-7-sulfo-2-heptenoic acid sodium salt; Z-2-( Z-2-(2,2-dimethylcyclopropanecarboxamide)-8-hydroxy-2-octenoic acid; Z-8-acetoxy -2-(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid. 2-8-cyano-2(2,2-dimethylcyclopropanecarboxamide)-2-octenoic acid. -(2,2-dimethylcyclopropane carboxamide)-2-
Octenoic acid and 100mg of NaCN in 2ml of DMSO
was produced by heating it to 80°C for 30 minutes. After extraction and purification 102 mg of colorless solid, mp 99-103°C, was collected. Analysis (C ₁₅ H ₂₂ N ₂ O ₃ ) Calculated value Actual value C 64.73 64.69 H 7.97 8.14 N 10.06 9.41

Claims (1)

Claims: 1. 6- and (optionally) 2-substituted pen-2- wherein the ratio of thienamycin type compound to dipeptidase inhibitor is in the range of about 1:3 to about 30:1.
Em-3-carboxylic acid, 1-carbadethia-pen-2-em-3-carboxylic acid or 1-azabicyclo(3.2.0.)hept-2-en-7-one-2
- Antibacterial compositions consisting of compounds of the thienamycin type, which are carboxylic acids, and dipeptidase (EC3.4.13.11) inhibitors. 2 The thienamycin compound is a compound of the following formula [wherein X can be _CH2 or S ^{; R2} is hydrogen; _-S - _CH2CH2NHR3 ( ^R3 is hydrogen, acetyl, formimidoyl, acetimidoyl); -S(O)-CH= _CHNHCOCH3 and -S-CH=
CHMHCOCH ₃ can be CHMHCOCH 3 and R ⁶ is [formula] (R ⁷ is hydrogen, hydroxyl or sulfonyloxy) or R ⁶ is H and all possible stereoisomers are as described above. Included in the structure definition. ] The composition according to claim 1. 3. The composition of claim 1, wherein the formulation is mixed with a pharmaceutical carrier. 4. The composition according to claim 3, in which a carrier is applied for injection. 5. The composition of claim 1, wherein the thienamycin type compound is thienamycin. 6 Claim 1 in which the thienamycin type compound is N-formimidoyl thienamycin
Compositions as described in Section. 7 Claim 1 in which the thienamycin type compound is N-acetimidoyl thienamycin
Compositions as described in Section. 8 Dipeptidase inhibitor is a compound of the following formula [In the formula, R ² and R ³ are hydrocarbon groups having 3 to 10 carbon atoms and 1 to 15 carbon atoms, respectively;
1 on either R ² or R ³ hydrocarbon chain
-6 hydrogens may be replaced by halogens or non-terminal methylenes may be replaced by oxygen or sulfur (including oxidized forms of sulfur);
Furthermore, R ³ and the terminal hydrogen can also be substituted with hydroxyl or thiol groups (which may be acylated or carbamoylated); or this hydrogen can be substituted with an amino group (which may be acylamino, ureido, amidino, guanidino, or alkyl,
or a derivative of a substituted amino group containing a quaternary nitrogen functional group);
or may alternatively be substituted by an acid group such as carboxyl, phosphate or sulfate group, or an ester or amide thereof, or cyano; or a combination thereof, such as a terminal amino acid group. and R ¹ is hydrogen, lower alkyl (C _1-6 ), dialkylaminoalkyl, or a pharmaceutically acceptable cation. 9 R ² is R ⁴ (R ⁴ is a molecule consisting of 3-10 carbon atoms or a cyclic hydrocarbon) provided that the carbon adjacent to the carbonyl is not quaternary; −R ⁵ R ⁶ [R ⁵ is a cyclic alkyl of 3-6 carbon atoms, R ⁶ is 1 to 2 alkyl substituents (which may be joined together to form another cycloalkyl group), or R ⁶ is 1 to 2 alkyl substituents (which may be joined together to form another cycloalkyl group); -R ⁷ R ⁸ (R ⁷ is alkylene of 1-3 carbon atoms and R ⁸ is cycloalkyl of 3-6 carbon atoms); or
9. The composition of claim 8, wherein ^R2 is straight chain, molecular, or alkylcycloalkyl and dialkylcycloalkyl of 3 to 10 carbon atoms. 9. The composition of claim 8, wherein 10 R ² is 2,2-dimethylcyclopropyl. 11. The composition according to claim 8, wherein ^R2 is 2,2-dichlorocyclopropyl. 12 The dipeptidase inhibitor is Z-2-(2,2
-dimethylcyclopropanecarboxamide)-2
The composition according to claim 8, which is octenoic acid.