Immunosuppression modulating compounds
The present invention provides immunosuppression compounds to inhibit the programmed cell death 1 (PD1) signalling pathway. The present invention further provides peptide based compositions for treatment of cancer or treatment of infections via immunopotentiation caused by inhibition of immunosuppressive signaling induced by PD-1, PD-L1, or PD-L2 and therapies using them, immunopotentiative substrates included as the active ingredient. Further, the invention provides an application of the compositions containing the peptide moieties for preventive and/or therapeutic agents for cancer, cancer metastasis, immunodeficiency, an infectious disease or the like and an application of peptide moieties as a testing or diagnostic agent or a research agent for such a disease.
1. A compound of formula (I)
or pharmaceutically acceptable salt thereof; wherein;
A is an amino acid sequence of from three amino acids to the full length of a peptide sequence of mammalian PD1 ectodomain fragments selected from FG loop, BC loop, C strand, or C′ strand to C′-C″ loop;
B is an amino acid sequence of from three amino acids to the full length of a peptide sequence of mammalian PD1 ectodomain fragments selected from BC loop, FG loop, C C′ loop to C′ strand, D strand, C′ strand to C′-C″ loop, or is absent;
with the proviso that if A is the amino acid sequence SNTSESF (SEQ ID NO: 4) and X is lysine then B cannot be the amino acid sequence SNTSESF SEQ ID NO: 4);
Z is the amino acid sequence FRVTQLAPKAQIKE (SEQ ID NO: 84);
D is up to two peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from BC loop, FG loop, or is absent;
E is up to four peptide sequences arranged in any order each being of from three amino acids up to the full length of a mammalian PD1 ectodomain fragment selected from D strand, FG loop, G strand, or is absent;
X and X′ are independently selected from lysine, ornithine, diaminopropionic acid, diaminobutyric acid or olefinic amino acid of formula
which is optionally linked with an additional lysine; or
X and X′ combine together forming a ring with olefinic amino acid which is optionally linked with an additional lysine; or
X′ is absent;
‘m’ is an integer selected from 1 to 6, both inclusive;
R 2 and R 3 are independently selected from group consisting of C 2 -C 20 acyl, absent or R a -L′;
R a is maleimido propionic acid;
L′ is —CO(CH 2 —CH 2 —O—) n NH; and
‘n’ is an integer selected from 2 to 10, both inclusive;
R 4 and R 5 are independently NH 2 , or one or both of R 4 or R 5 are absent, with the proviso to the compound of Formula (I), that in a compound of Formula (I) as above defined:
a) up to 5 but not more than 25% of the amino acids may be substituted with other natural or unnatural amino acids;
b) not more than 30% of the amino acids may be omitted;
c) in each said peptide sequence up to 2 amino acids may be added individually at any position;
d) up to 5 but not more than 25% of the peptide bonds may instead be replaced by reduced amide bond (—CH 2 NH—);
e) up to 100% of the amino acids may be D-amino acids;
f) up to 100% of the amino acids may be in reverse order.
2. The compound according to claim 1 , wherein said mammalian PD1 ectodomain fragments are from human, murine, dog, horse or rat PD1.
3. The compound according to claim 1 , wherein;
A is an amino acid sequence of from three amino acids to the full length of a peptide sequence of human or murine PD1 ectodomain fragments selected from FG loop, BC loop, C strand, or C′ strand to C′-C″ loop as in table below;
hPD-1
mPD-1
BC Loop
SNTSESF
SNWSEDL
(SEQ ID NO: 4)
(SEQ ID NO: 27)
C Strand
VLNWYRM
MLNWNRL
(SEQ ID NO: 5)
(SEQ ID NO: 28)
C′ strand to
FPED
FCNG
C′-C″ loop
(SEQ ID NO: 21)
(SEQ ID NO: 37)
FG loop
LAPKA
LHPKA
(SEQ ID NO: 15)
(SEQ ID NO: 35)
B is an amino acid sequence of from three amino acids to the full length of a peptide sequence of human or murine PD1 ectodomain fragments selected from BC loop, FG loop, C C′ loop to C′ strand, D strand, C′ strand to C′-C″ loop as in table below, or is absent;
hPD-1
mPD-1
BC Loop
SNTSESF
SNWSEDL
(SEQ ID NO: 4)
(SEQ ID NO: 27)
C′ strand to
FPED
FCNG
C′-C″ loop
(SEQ ID NO: 21)
(SEQ ID NO: 37)
C C′ loop to
SPSNQTDKLAAFP
SPSNQTEKQAAFC
C′ strand
(SEQ ID NO: 23)
(SEQ ID NO: 39)
D Strand
FRVTQ
FQIIQ
(SEQ ID NO: 11)
(SEQ ID NO: 33)
FG loop
LAPKA
LHPKA
(SEQ ID NO: 15)
(SEQ ID NO: 35)
with the proviso that is A is the amino acid sequence SNTSESF (SEQ ID NO: 4) and X is lysine then B cannot be amino acid sequence SNTSESF (SEQ ID NO: 4);
Z is the amino acid sequence FRVTQLAPKAQIKE (SEQ ID NO: 84);
D is up to two peptide sequences arranged in any order each being of from three amino acids up to the full length of a human or murine PD1 ectodomain fragment selected from BC loop, FG loop, as in table below or is absent;
hPD-1
mPD-1
BC Loop
SNTSESF
SNWSEDL
(SEQ ID NO: 4)
(SEQ ID NO: 27)
FG loop
LAPKA
LHPKA
(SEQ ID NO: 15)
(SEQ ID NO: 35)
E is up to four peptide sequences arranged in any order each being of from three amino acids up to the full length of a human or murine PD1 ectodomain fragment selected from D strand, FG loop, G strand, or is absent;
hPD-1
mPD-1
D Strand
FRVTQ
FQIIQ
(SEQ ID NO: 11)
(SEQ ID NO: 33)
FG loop
LAPKA
LHPKA
(SEQ ID NO: 15)
(SEQ ID NO: 35)
G Strand
QIKE
KIEE
(SEQ ID NO: 16)
(SEQ ID NO: 36)
X and X′ are independently selected from lysine, ornithine, diaminopropionic acid, diaminobutyric acid or olefinic amino acid of formula
which is optionally linked with an additional lysine; or
X and X′ combine together forming a ring with olefinic amino acid which is optionally linked with an additional lysine; or
X′ is absent;
‘m’ is an integer selected from 1 to 6, both inclusive;
R 2 and R 3 are independently selected from group consisting of C 2 -C 20 acyl, absent or R a -L′;
R a is maleimido propionic acid;
L′ is —CO(CH 2 —CH 2 —O—) n NH—; and
‘n’ is an integer selected from 2 to 10, both inclusive;
R 4 and R 5 are independently NH 2 , or one or both of R 4 or R 5 are absent, with the proviso to the compound of Formula I, that in a compound of Formula I as above defined:
a) up to 5 but not more than 25% of the amino acids may be substituted with other natural or unnatural amino acids;
b) not more than 30% of the amino acids may be omitted;
c) in each said peptide sequence up to 2 amino acids may be added individually at any position;
d) up to 5 but not more than 25% of the peptide bonds may instead be replaced by reduced amide bond (—CH 2 NH-);
e) up to 100% of the amino acids may be D-amino acids;
f) up to 100% of the amino acids may be in reverse order.
4. The compound according to claim 1 , wherein X is lysine and X′ is absent.
5. The compound according to claim 1 , wherein both X and X′ are lysine.
6. The compound according to claim 1 , wherein R 2 and R 3 are selected from C 2 -C 20 acyl or R a -L′; wherein,
R a is maleimido propionic acid;
L′ is CO(CH 2 —CH 2 —O—) n NH—; and
‘n’ is an integer selected from 2 to 10, both inclusive.
7. The compound according to claim 1 , wherein X′, D, E and R 5 are absent and R 4 is NH 2 .
8. A compound selected from
Comp.
Sequence
No.
Compound
ID
6
FRVTQK(SNTSESF)FRVTQAhxLAPKAQIKE-NH2
47
7
FRVTQK(SNTSESF)FRVTQLAPKAQIKE-NH2
48
11
SNTSESFK(SNQTDK)FRVTQLAPKAQIKE-NH2
52
12
SNTSESFK(SNQTDK)FRVTQK(LAP)AQIKE-NH2
53
22
63
23
64
24
65
27
SNTSESFKFRVTQLAPKAQIKE-NH2
68
28
SNTSESFKSNTSESFFRVTQLAPKAQIKE-NH2
69
37
SNTSESFK(LAP)FRVTQLAPKAQIKE-NH2
78
38
LAPK(LAP)FRVTQLAPKAQIKE-NH2
79
39
LAPKAQIKE-NH2
80
48
EKIQAKPALQTVRFK(FSESTNS)FSESTNS-NH2
89
49
ekiqakpalqtvrfk(fsestns)fsestns-NH 2 Retro inverso
90
(All D-amino acid)
69
SNWSEDLK(SNWSEDL)FQIIQLHPKAKIEE-NH2
110
70
EDK(ED)FRVTQLAPKAQIKE-NH2
111
71
SNTSESFK(ED)FRVTQLAPKAQIKE-NH2
112
72
LAPKAK(ED)FRVTQLAPKAQIKE-NH2
113
73
SNQTDKK(ED)FRVTQLAPKAQIKE-NH2
114
75
SNQTDKK(VLNWYRM)GIYLCGAISLAPKAQIKE-NH2
116
76
EDK(VLNWYRM)GIYLCGAISLAPKAQIKE-NH2
117
79
VLNWYRM(SNQTDK)GIYLCGAISLAPKAQIKE-NH2
120
82
SNWSEDLK(SNWSEDL)FQIIQLHPK(CH 3 (CH 2 ) 14 CO)AKIEE-NH2
123
91
SNWSEDLK(SNWSEDL)FQIIQLHPK(MPA—NH—CH 2 —CH 2 —O—CH 2 —CH 2 —O—CO)AKIEE-
132
NH2
99
SNTSESF-Orn-(SNTSESF)KAQIKE-NH2
140
100
SNTSESF-Dap-(SNTSESF)KAQIKE-NH2
141
101
SNTSESF-Dab-(SNTSESF)KAQIKE-NH2
142
107
LAPKA(SNQTDK)FRVTQLAPKAQIKE-NH2
148.
9. A compound according to claim 3 having formula (Ia)
or pharmaceutically acceptable salt thereof;
wherein;
A is an amino acid sequence of from three amino acids to the full length of a peptide sequence of human or murine PD1 ectodomain fragments selected from FG loop, BC loop, C strand, or C′ strand to C′-C″ loop;
B is an amino acid sequence of from three amino acids to the full length of a peptide sequence of human or murine PD1 ectodomain fragments selected from BC loop, FG loop, C-C′ loop to C′ strand, D strand, C′ strand to C′-C″ loop or is absent;
with the proviso that if A is the amino acid sequence SNTSESF (SEQ ID NO: 4) and X is lysine then B cannot be the amino acid sequence SNTSESF SEQ ID NO: 4);
Z is the amino acid sequence FRVTQLAPKAQIKE (SEQ ID NO: 84); and
X is lysine.
10. The compound according to claim 9 , wherein A and B are independently an amino acid sequence of from three amino acids to the full length of said BC loop, with the proviso that if A is the amino acid sequence SNTSESF (SEQ ID NO: 4) and X is lysine then B cannot be the amino acid sequence SNTSESF (SEQ ID NO: 4).
11. The compound according to claim 9 , wherein R 2 and R 3 are absent.
12. The compound according to claim 9 , wherein R 3 is C16-acyl.
13. The compound according to claim 9 , wherein up to three amino acids are D-amino acids.
14. The compound according to claim 9 , wherein all amino acids are D-amino acids.
15. The compound according to claim 9 , comprising up to three reduced amide bonds (—CH 2 NH—).
16. A pharmaceutical composition comprising a compound of formula (I) or a pharmaceutically acceptable salt thereof as claimed in claim 1 , and a pharmaceutically acceptable diluent or carrier.
17. A method of modulating an immune response mediated by PD-1 signaling pathway in a subject, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1 , such that the immune response in the subject is modulated.
18. A method of inhibiting growth of tumor cells and/or metastasis in a subject, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1 capable of inhibiting the programmed cell death 1 (PD1) signaling pathway.
19. The method of claim 18 , wherein the tumor cells are of a cancer selected from the group consisting of melanoma, renal cancer, prostate cancer, breast cancer, colon cancer, and lung cancer.
20. The method of claim 19 , wherein the tumor cells are of a cancer selected from bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, cutaneous or intraocular malignant melanoma, uterine cancer, ovarian cancer, rectal cancer, cancer of the anal region, stomach cancer, testicular cancer, uterine cancer, carcinoma of the fallopian tubes, carcinoma of the endometrium, carcinoma of the cervix, carcinoma of the vagina, carcinoma of the vulva, Hodgkin's Disease, non-Hodgkin's lymphoma, cancer of the esophagus, cancer of the small intestine, cancer of the endocrine system, cancer of the thyroid gland, cancer of the parathyroid gland, cancer of the adrenal gland, sarcoma of soft tissue, cancer of the urethra, cancer of the penis, chronic or acute leukemias including acute myeloid leukemia, chronic myeloid leukemia, acute lymphoblastic leukemia, chronic lymphocytic leukemia, solid tumors of childhood, lymphocytic lymphoma, cancer of the bladder, cancer of the kidney or ureter, carcinoma of the renal pelvis, neoplasm of the central nervous system (CNS), primary CNS lymphoma, tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma, Kaposi's sarcoma, epidermoid cancer, squamous cell cancer, T-cell lymphoma, environmentally induced cancers including those induced by asbestos, and combinations of said cancers.
21. A method of treating an infectious disease in a subject, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1 to inhibit the programmed cell death 1 (PD1) signaling pathway such that the subject is treated for the infectious disease.
22. A method of treating bacterial and viral infections in a subject, comprising administering to the subject a therapeutically effective amount of the compound according to claim 1 to inhibit the programmed cell death 1 (PD1) signaling pathway such that the subject is treated for the bacterial and viral infections.