US 2017/0096692 A1 · CELL-FREE PREPARATION OF CARBAPENEMS

Abstract

Provided herein are cell-free systems for generating carbapenems, e.g., a compound of the Formula (I): or salts thereof; wherein , R1, R2, R3, R4, R5 and R6 are defined herein. Also provided are pharmaceutical compositions comprising a compound generated by the inventive cell-free system, and use of these compounds and compositions for the treatment of bacterial infections.

Claims25

Dependent claims

1-32. (canceled)

One or more bacterial cells, wherein each cell expresses one or more enzymes selected from the group consisting of:

a. ProA and ProB, or γ-glutamyl kinase-GP-reductase multi-enzyme complex;
b. Glk;
c. Pgi;
d. PfkA;
e. Fba;
f. TpiA;
g. GapA;
h. Pgk;
i. GpmA;
j. Eno;
k. PykA;
l. PykF;
m. AceE;
n. AceF;
o. acetyl-CoA acetyltransferase;
p. S-hydratase and 3-hydroxyacyl-CoA dehydrogenase, or beta-oxidation multi-enzyme;
q. crotonyl-CoA reductase;
r. carboxymethyl-Pro synthase;
s. carbapenam synthetase, or beta-lactam synthetase;
t. carbapenem synthase;
and isozymes thereof;
wherein the one or more cells, in combination, express all enzymes (a)-(t), or isozyme(s) thereof;
and
wherein a lysate of the one or more cells is capable, upon contact with glucose and an optionally substituted glutamate of the formula (i):
or salt thereof,
of generating a carbapenem of Formula (I-a):
or a salt or tautomer thereof,
wherein:
the dashed line represents a double bond;
R1 is hydrogen;
R2 s —CH2CH3;
each instance of R3 and R4 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
R5 is hydrogen or —OR8;
each instance of R8 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)R8a, —C(═O)OR8a, —C(═O)SR8a, —C(═O)N(R8b)2, —C(═NR8b)R8a, —C(═NR8b)OR8a, —C(═NR8b)SR8a, —C(═NR8b)N(R8b)2, —C(═S)R8a, —C(═S)OR8a, —C(═S)SR8a, —C(═S)N(R8b)2, —C(═O)NR8bSO2R8a, —S(═O)R8a, —SO2R8a, —SO2N(R8a)2, —Si(R8a)3, —P(O)(R8a)2, —P(═O)(OR8a)2, —P(═O)(R8a)(OR8a), —P(═O)(R8a)(N(R8b)2), —P(═O)(N(R8b)2)2, —P(═O)2R8a, —P(═O)2OR8a, —P(═O)2N(R8b)2, —B(R8a)2, —B(OR8a)2, and —BR8a(OR8a);
each instance of R8a is independently selected from the group consisting of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, if attached to an oxygen atom an oxygen protecting group, and if attached to a sulfur atom a sulfur protecting group;
each instance of R8b is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, and a nitrogen protecting group, or two R8b are joined to form an optionally substituted heterocyclic ring;
or two R8a groups or an R8a and R8b group are joined to form an optionally substituted heterocyclic ring;
and
R7a is hydrogen.

36 more elements

One or more bacterial cells, wherein each cell expresses one or more enzymes selected from the group consisting of:

a. ProA and ProB, or γ-glutamyl kinase-GP-reductase multi-enzyme complex;
b. Glk;
c. Pgi;
d. PfkA;
e. Fba;
f. TpiA;
g. GapA;
h. Pgk;
i. GpmA;
j. Eno;
k. PykA;
l. PykF;
m. AceE;
n. AceF;
o. acetyl-CoA carboxylase;
p. carboxymethyl-Pro synthase;
q. carbapenam synthetase, or beta-lactam synthetase;
r. carbapenem synthase;
and isozymes thereof;
wherein the one or more cells, in combination, express all enzymes (a)-(r), or isozyme(s) thereof;
and
wherein a lysate of the one or more cells is capable, upon contact with glucose and an optionally substituted glutamate of the formula (i):
or salt thereof, —of generating a carbapenem of Formula (I-a):
or a salt or tautomer thereof,
wherein:
the dashed line represents a double bond;
R1 is hydrogen;
R2 is hydrogen;
each instance of R3 and R4 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
R5 is hydrogen or —OR8;
each instance of R8 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)R8a, —C(═O)OR8a, —C(═O)SR8a, —C(═O)N(R8b)2, —C(═NR8b)R8a, —C(═NR8b)OR8a, —C(═NR8b)SR8a, —C(═NR8b)N(R8b)2, —C(═S)R8a, —C(═S)OR8a, —C(═S)SR8a, —C(═S)N(R8b)2, —C(═O)NR8bSO2R8a, —S(═O)R8a, —SO2R8a, —SO2N(R8a)2, —Si(R8a)3, —P(═O)(R8a)2, —P(═O)(OR8a)2, —P(═O)(R8a)(OR8a), —P(═O)(R8a)(N(R8b)2), —P(═O)(N(R8b)2)2, —P(═O)2R8a, —P(═O)2OR8a, —P(═O)2N(R8b)2, —B(R8a)2, —B(OR8a)2, and —BR8a(OR8a);
each instance of R8a is independently selected from the group consisting of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, if attached to an oxygen atom an oxygen protecting group, and if attached to a sulfur atom a sulfur protecting group;
each instance of R8b is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, and a nitrogen protecting group, or two R8b are joined to form an optionally substituted heterocyclic ring;
or two R8a groups or an R8a and R8b group are joined to form an optionally substituted heterocyclic ring;
and
R7a is hydrogen.

33 more elements

Dependencies are inferred from the claim text.

Facts

Application

15/291,943Filed 12 Oct 2016

Priority

9 Sep 2011

Published

6 Apr 2017

Term ends

Not estimatedThe source records don’t support an estimate.

Classification22 codes

C12P 17/184
Heterocyclic compounds containing nitrogen atoms as the only ring heteroatoms in the condensed system: containing a beta-lactam ring, e.g. thienamycin
A61P 31/04
Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics: antibacterial agents
C12N 9/0008
Oxidoreductases: acting on the aldehyde or oxo group of donors
C12N 9/0026
Acting on nitrogen containing compounds as donors: acting on CH-NH groups of donors

Family

Application61/533,039Us provisional application · filed 9 Sep 2011

Application13/606,911Continuation · filed 7 Sep 2012

Application15/291,943Continuation

US 2013/0065878 A1Published application

US 9,469,861 B2Related patent

US 2017/0096692 A1This patent

From the US filing history. Foreign family members aren’t listed.

Abstract

Claims25

Dependent claims

1-32. (canceled)

One or more bacterial cells, wherein each cell expresses one or more enzymes selected from the group consisting of:

a. ProA and ProB, or γ-glutamyl kinase-GP-reductase multi-enzyme complex;
b. Glk;
c. Pgi;
d. PfkA;
e. Fba;
f. TpiA;
g. GapA;
h. Pgk;
i. GpmA;
j. Eno;
k. PykA;
l. PykF;
m. AceE;
n. AceF;
o. acetyl-CoA acetyltransferase;
p. S-hydratase and 3-hydroxyacyl-CoA dehydrogenase, or beta-oxidation multi-enzyme;
q. crotonyl-CoA reductase;
r. carboxymethyl-Pro synthase;
s. carbapenam synthetase, or beta-lactam synthetase;
t. carbapenem synthase;
and isozymes thereof;
wherein the one or more cells, in combination, express all enzymes (a)-(t), or isozyme(s) thereof;
and
wherein a lysate of the one or more cells is capable, upon contact with glucose and an optionally substituted glutamate of the formula (i):
or salt thereof,
of generating a carbapenem of Formula (I-a):
or a salt or tautomer thereof,
wherein:
the dashed line represents a double bond;
R1 is hydrogen;
R2 s —CH2CH3;
each instance of R3 and R4 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
R5 is hydrogen or —OR8;
each instance of R8 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)R8a, —C(═O)OR8a, —C(═O)SR8a, —C(═O)N(R8b)2, —C(═NR8b)R8a, —C(═NR8b)OR8a, —C(═NR8b)SR8a, —C(═NR8b)N(R8b)2, —C(═S)R8a, —C(═S)OR8a, —C(═S)SR8a, —C(═S)N(R8b)2, —C(═O)NR8bSO2R8a, —S(═O)R8a, —SO2R8a, —SO2N(R8a)2, —Si(R8a)3, —P(O)(R8a)2, —P(═O)(OR8a)2, —P(═O)(R8a)(OR8a), —P(═O)(R8a)(N(R8b)2), —P(═O)(N(R8b)2)2, —P(═O)2R8a, —P(═O)2OR8a, —P(═O)2N(R8b)2, —B(R8a)2, —B(OR8a)2, and —BR8a(OR8a);
each instance of R8a is independently selected from the group consisting of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, if attached to an oxygen atom an oxygen protecting group, and if attached to a sulfur atom a sulfur protecting group;
each instance of R8b is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, and a nitrogen protecting group, or two R8b are joined to form an optionally substituted heterocyclic ring;
or two R8a groups or an R8a and R8b group are joined to form an optionally substituted heterocyclic ring;
and
R7a is hydrogen.

36 more elements

One or more bacterial cells, wherein each cell expresses one or more enzymes selected from the group consisting of:

a. ProA and ProB, or γ-glutamyl kinase-GP-reductase multi-enzyme complex;
b. Glk;
c. Pgi;
d. PfkA;
e. Fba;
f. TpiA;
g. GapA;
h. Pgk;
i. GpmA;
j. Eno;
k. PykA;
l. PykF;
m. AceE;
n. AceF;
o. acetyl-CoA carboxylase;
p. carboxymethyl-Pro synthase;
q. carbapenam synthetase, or beta-lactam synthetase;
r. carbapenem synthase;
and isozymes thereof;
wherein the one or more cells, in combination, express all enzymes (a)-(r), or isozyme(s) thereof;
and
wherein a lysate of the one or more cells is capable, upon contact with glucose and an optionally substituted glutamate of the formula (i):
or salt thereof, —of generating a carbapenem of Formula (I-a):
or a salt or tautomer thereof,
wherein:
the dashed line represents a double bond;
R1 is hydrogen;
R2 is hydrogen;
each instance of R3 and R4 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, and optionally substituted heteroaryl;
R5 is hydrogen or —OR8;
each instance of R8 is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, —C(═O)R8a, —C(═O)OR8a, —C(═O)SR8a, —C(═O)N(R8b)2, —C(═NR8b)R8a, —C(═NR8b)OR8a, —C(═NR8b)SR8a, —C(═NR8b)N(R8b)2, —C(═S)R8a, —C(═S)OR8a, —C(═S)SR8a, —C(═S)N(R8b)2, —C(═O)NR8bSO2R8a, —S(═O)R8a, —SO2R8a, —SO2N(R8a)2, —Si(R8a)3, —P(═O)(R8a)2, —P(═O)(OR8a)2, —P(═O)(R8a)(OR8a), —P(═O)(R8a)(N(R8b)2), —P(═O)(N(R8b)2)2, —P(═O)2R8a, —P(═O)2OR8a, —P(═O)2N(R8b)2, —B(R8a)2, —B(OR8a)2, and —BR8a(OR8a);
each instance of R8a is independently selected from the group consisting of optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, if attached to an oxygen atom an oxygen protecting group, and if attached to a sulfur atom a sulfur protecting group;
each instance of R8b is independently selected from the group consisting of hydrogen, optionally substituted alkyl, optionally substituted alkenyl, optionally substituted alkynyl, optionally substituted heteroalkyl, optionally substituted heteroalkenyl, optionally substituted heteroalkynyl, optionally substituted carbocyclyl, optionally substituted heterocyclyl, optionally substituted aryl, optionally substituted heteroaryl, and a nitrogen protecting group, or two R8b are joined to form an optionally substituted heterocyclic ring;
or two R8a groups or an R8a and R8b group are joined to form an optionally substituted heterocyclic ring;
and
R7a is hydrogen.

33 more elements

Dependencies are inferred from the claim text.

Cell-free preparation of carbapenems

Life of this patent

Abstract

Claims25

Cell-free preparation of carbapenems

Life of this patent

Abstract

Claims25