Medicinal Chemistry

Cilastatin metabolism led drug discovery

Beta Lactams Cell Wall synthesis inhibitors

Penicillin G

Piperacillin

Ceftazidime

Aztreonam

Beta Lactams Cell Wall synthesis inhibitors

Imipenem

Panipenem

Doripenem

Ertapenem

Meropenem

Faropenem

Sanfitrinem

Imipenem
H HO H S NH 2 H 3C O O N OH

During the development of carbapenems, imipenem (Nformidoyl derivative of thienamycin) had the highest potency, broad spectrum activity and lack of cross resistance. Disadvantages: Low level of urinary concentrations of the antibiotic due to extensive renal metabolism; nephrotoxicity

Thienamycin antibiotic
H HO H S N H H3 C O O N OH NH

Imipenem antibiotic

Human Renal dehydropeptidase

Membrane bound glycoprotein involved in hydrolysis of dipeptides located in renal cortex Metallo enzyme (Zinc cofactor) Metabolism of glutathione its conjugates and some drugs eg. Imipenem, Panipenem To date the only mammalian enzyme able to hydrolyze the beta-lactam ring Dehydropeptidase-I is responsible for hydrolyzing the beta lactam ring of imipenem, inactivating it but does not affect penicillins or cephalosporins

S HO O HN H 2N COOH

Cilastatin

Cilastatin was developed as a reversible, competitive inhibitor of dehydropeptidase I (DHP-I) on the basis of the structural similarities between the scissile bonds in imipenem and dehydropeptides In the presence of cilastatin, dehydropeptidase does not open the lactam ring of imipenem (Clinically it is given as 1 : 1 combination) Affinity for cilastatin is 30000 times greater than imipenem for DHP-I enzyme Cilastatin prevents renal re-absorption of imipenem and increases its excretion in urine, thereby imipenems usage is now broadened to indications such as urinary tract infection which was otherwise limited to only systemic infections. Because in the absence of cilastatin, required concentration of of active drug of imipenem in urine was not feasible since dehydropeptidase metabolises imipenem in kidney.

COOH O HN NH2

glycyl dehydrophenylalanine A Substrate for Dehydropeptidase

COOH O HN

COOH

R1 (Z)-2-benzamidobut-2-enoic acid Lead

HN

(Z)-2-(Acylamino)-2-butenoic acids Activity

H COOH O HN
Ki M (% inhibition at 100M)
(30)

Contd

R
Ki M (% inhibition at 100M)
70
Cl

Ki M (% inhibition at 100M)
(7)

(2.5)

(3)

(3)

(0) (21)
MeO S

10

2-benzamido-2-butenoic acid as a lead, a number of other aryl and heteroaryl groups were tested as replacements for phenyl with unimpressive results.

(Z)-2-(Acylamino)-2-butenoic acids - Activity

COOH O

Ki M (% inhibition at 100M) (12) (43) 10

HN

R
(CH 2)4 CH3

Ki M (% inhibition at 100M) (54)

CH(Me)CH(CH 3) 2

Ki M (% inhibition at 100M) 3.3

R=

Me Et

(CH 2)5 CH3

(49)

CH(iPr)2

(13)

(CH 2)9 CH3 (CH 2)10 CH 3

19 35

(CH 2)2 CH(CH 3) 2

14

(CH 2)2 CH3

30 32

(CH 2)4 CH(CH 3) 2

(57)

(CH 2)3 CH3

CH 2CH(CH 3) 2

CH=C(CH3 )2

28

(Z)-2-(Acylamino)-2-butenoic acids Activity

Contd
H COOH O HN
Ki M (% inhibition at 100M)

Ki M (% inhibition at 100M)

22

30

9.8

15

25

(3)

(33)

Other than the cyclopropyl shown above larger cycloalkyl groups exerted less activity

(Z)-2-(Acylamino)-2-butenoic acids Activity

H COOH O HN
Ki M (% inhibition at 100M)
(16) 4.6 0.4 1.7
Cl

Contd

R
Ki M (% inhibition at 100M)

Ki M (% inhibition at 100M)

Ki M (% inhibition at 100M)

0.08

( )
7.2

Cl

0.19 12

Br

0.03

(+)
1.6 19.8

Br

(-)
6.5 1.6

(10)

(11)

COOH O

HN

R=

H Et

52 0.18 10

(CH 2 )4 CH 3

8.8
3 N

3.45

(CH 2 )7 CH 3

0.11
4
N N

0.84

(CH 2 )2 CH=CH 2 CH 2 CF3

0.14 0.24 0.21 0.23 1 0.048

0.092
(CH 2 )4 CN (CH 2 )5 OH (CH 2 )2 CH 3 4 N

1.10

0.11
(CH 2 )5 NH 2

4 N

OH

0.91

(CH 2 )3 CH 3

0.11
(CH 2 )3 COOH

3S

P O

OH OH

4 N

1.09

0.22

JMC 1987 30 1074

COOH O HN
O

OH S NH2 HN

O OO Na +

Cilastatin 0.11

R=
3S O OH O ONa O

0.087
3S
COOH

0.04

2S O

OMe

0.28

0.56
N

3N

0.13

2S NH 2 O 3S

OH

0.27

3S
(CH 2) 2COOH

0.14

OH

OH NH 2 O

0.21

4N H

P O

OH OH

0.4

4S

CONH2

0.25
3S

0.19
OH

4N H

P O

OH OH

0.28

5N O

OH

0.4

NH 2

Compared with the basic amines, acidic groups such phosphates and carboxylates showed better activity

JMC 1987 30 1074

Highlights - SAR

COOH
HN

O OO Na + O

OH S NH2 HN

O OO Na +

R1

HN O

Compd 1

Cilastatin

SAR study reveals that a large number of diverse substituents can be tolerated in R1 of the 2-(2,2-dimethylcyclopropylamino)-2-butenoic acid derivatives ,- unsaturated acid moiety and the allylic CH2 of the R1 chain are important for the activity Compd 1 and cilastatin showed desired pharmacokinetic properties, wherein Compd 1 caused local irritation when injected repeatedly at high doses in animals N-acetyl cilastatin is the active metabolite found in body which is also twice as active as cilastatin. But half life of N-acetyl cilastatin is lesser than cilastatin and hence it could not be profiled for further studies

NH2

S COOEt

Br(CH2)5Br

NaH DMF Toluene

Br S S

NBS, ACN
Br

HBr / HOAc
O Br

5 COOEt

5 COOEt
H S COOH

5 COOH

Toluene PTSA (cat.) ref lux

H Br

COOH

NaOH
O HS NH 2 OH H2 N

HN C O

4
O

HN C O

OH

Z)-7-[(2R)-2-amino-3-hydroxy-3-oxopropyl]sulfanyl-2-{[(1S)2,2-dimethylcyclopropanecarbonyl]amino}hept-2-enoic acid

EP0048301 & WO 03018544