1. A method of determining inhibitory activity of a compound of formula (I), the method
comprising the steps of:
(i) obtaining a biological sample and determining expression level of TIGIT and at
least one biomarker selected from the group consisting of PD-L1 and PD-L2;
(ii) comparing the expression levels in step (i) with control samples, independently, to
determine overexpression of TIGIT and the biomarker in the biological sample; and
(iii)contacting the biological sample with a compound of formula (I)
or a stereoisomer thereof or a pharmaceutically acceptable salt thereof to determine
inhibitory activity of the compound;
wherein the inhibitory activity of the compound is measured by assessing
modulation of TIGIT signaling pathway and PD-1/PD-L1 or PD-L2 signaling
pathway,
wherein,
R1 represents hydrogen, -(C1-C6) alkyl optionally substituted with -OH, -COOH,
aryl, heteroaryl, or aryl-OH;
R2 represents hydrogen, -(C1-C6) alkyl optionally substituted with -OH, -SH,
C(O)NH2, -COOH, aryl, heteroaryl, or aryl-OH;
Ra represents hydrogen; or Ra and R2 taken together with the atom to which they are
attached form a pyrrolidine ring;
R3 represents hydrogen or a group represented by formula (I’),
84
(I’);
----- represents point of attachment;
Rb represents hydrogen; or Rb and Rc taken together with the atoms to which they are
attached form a pyrrolidine ring; and
Rc represents hydrogen, -(C1-C6)alkyl optionally substituted with -OH, -C(O)NH2,
COOH, aryl, or aryl-OH.
2. The method as claimed in claim 1, wherein the compound is represented by compound
of formula (IA),
;
wherein, R1, R2, Ra, Rb, and Rc are as defined in claim 1.
3. The method as claimed in claim 1, wherein
R1 represents -(C1-C6)alkyl optionally substituted with -OH, -COOH, imidazolyl,
phenyl, or (p-OH)phenyl;
R2 represents hydrogen, -(C1-C6)alkyl substituted with -OH, -SH, -COOH, phenyl,
imidazolyl, or (p-OH)phenyl;
Ra represents hydrogen; or Ra and R2 taken together with the atom to which they are
attached form a pyrrolidine ring;
Rb represents hydrogen; or Rb and Rc taken together with the atoms to which they are
attached form a pyrrolidine ring; and
85
Rc represents hydrogen, -(C1-C6)alkyl optionally substituted with -OH, -C(O)NH2, -
COOH, phenyl, or (p-OH)phenyl.
4. The method as claimed in any one of claims 1 to 3, wherein the compound is
Compound No. Structure
1
;
2
;
3
;
4
;
5
;
6
;
86
Compound No. Structure
7
;
8
;
9
;
10
;
11
;
12
;
13
;
14
;
87
Compound No. Structure
15
;
16
;
17
;
18
;
19
; or
20
;
or a pharmaceutically acceptable salt or a stereoisomer thereof.
5. The method as claimed in claim 1, wherein the biological sample is selected from whole
blood, plasma, serum, cells (e.g., tumor cells), saliva, urine, stool and tissue.
88
6. The method as claimed in claims 1 to 5, wherein the overexpression of TIGIT, PD-L1
and PD-L2 is observed in disease or disorder selected from cancer, immune disorders,
immunodeficiency disorders, inflammatory disorders, infectious diseases, and
transplant rejection.
7. The method as claimed in claim 6, wherein the cancer is selected from blastoma, breast
cancer, epithelial cancer, colon cancer, lung cancer, melanoma, prostate cancer, renal
cancer, bone cancer, pancreatic cancer, skin cancer, cancer of the head or neck, uterine
cancer, ovarian cancer, colorectal cancer, rectal cancer, cancer of the anal region, cancer
of the peritoneum, stomach cancer, testicular cancer, carcinoma of the fallopian tubes,
carcinoma of the endometrium, cervical cancer, vaginal cancer, vulval cancer, 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, cancer of the urethra, cancer of the penis, solid tumors of childhood,
Hodgkin’s lymphoma, non-Hodgkin’s lymphoma, cutaneous T cell lymphoma,
mesothelioma, thymic carcinoma, myeloma, cancer of the bladder, cancer of the ureter,
carcinoma of the renal pelvis, liver cancer, pancreatic cancer, post-transplant
lymphoproliferative disorder (PTLD), neoplasm of the central nervous system (CNS),
tumor angiogenesis, spinal axis tumor, brain stem glioma, pituitary adenoma,
epidermoid cancer, salivary gland carcinoma, squamous cell cancer, abnormal vascular
proliferation associated with phakomatoses, edema (such as that associated with brain
tumors), Meigs’ syndrome, Merkel cell carcinoma, and environmentally induced
cancers.
8. The method as claimed in claim 6, wherein the infectious disease is selected from at
least one bacterium selected from anthrax, Bacilli, Bordetella, Borrelia, botulism,
Brucella, Burkholderia, Campylobacter, Chlamydia, cholera, Clostridium, Conococcus,
Corynebacterium, diptheria, Enterobacter, Enterococcus, Erwinia, Escherichia,
Francisella, Haemophilus, Heliobacter, Klebsiella, Legionella, Leptospira,
leptospirosis, Listeria, Lyme’s disease, meningococcus, Mycobacterium, Mycoplasma,
Neisseria, Pasteurella, Pelobacter, plague, Pneumonococcus, Proteus, Pseudomonas,
Rickettsia, Salmonella, Serratia, Shigella, Staphylococcus, Streptococcus, tetanus,
Treponema, Vibrio, Yersinia, and Xanthomonas; at least one virus selected from
arboviral encephalitis virus, adenovirus, herpes simplex type I, herpes simplex type 2,
89
Varicella-zoster virus, Epstein-barr virus, cytomegalovirus, herpesvirus type 8,
papillomavirus, BK virus, coronavirus, echovirus, JC virus, smallpox, Hepatitis B,
bocavirus, parvovirus B19, astrovirus, Norwalk virus, coxsackievirus, Hepatitis A,
poliovirus, rhinovirus, severe acute respiratory syndrome virus, Hepatitis C, yellow
fever, dengue virus, West Nile virus, rubella, Hepatitis E, human immunodeficiency
virus (HIV), human T-cell lymphotropic virus (HTLV), influenza, guanarito virus,
Junin virus, Lassa virus, Machupo virus, Sabia virus, Crimean- Congo hemorrhagic
fever virus, ebola virus, Marburg virus, measles virus, molluscum virus, mumps virus,
parainfluenza, respiratory syncytial virus, human metapneumovirus, Hendra virus,
Nipah virus, rabies, Hepatitis D, rotavirus, orbivirus, coltivirus, vaccinia virus, and
Banna virus; a fungal infection selected from thrush, Aspergillus (fumigatus, niger,
etc.), Blastomyces dermatitidis, Candida (albicans, krusei, glabrata, tropicalis, etc.),
Coccidioides immitis, Cryptococcus (neoformans, etc.), Histoplasma capsulatum,
Mucorales (mucor, absidia, rhizophus), Paracoccidioides brasiliensis,, sporotrichosis,
Sporothrix schenkii, zygomycosis, chromoblastomycosis, lobomycosis, mycetoma,
onychomycosis, piedra pityriasis versicolor, tinea barbae, tinea capitis, tinea corporis,
tinea cruris, tinea favosa, tinea nigra, tinea pedis, otomycosis, phaeohyphomycosis, and
rhinosporidiosis; and at least one parasite selected from Acanthamoeba, Babesia
microti, Balantidium coli, Entamoeba hystolytica, Giardia lamblia, Cryptosporidium
muris, Trypanosomatida gambiense, Trypanosomatida rhodesiense, Trypanosoma
brucei, Trypanosoma cruzi, Leishmania mexicana, Leishmania braziliensis,
Leishmania tropica, Leishmania donovani, Toxoplasma gondii, Plasmodium vivax,
Plasmodium ovale, Plasmodium malariae, Plasmodium falciparum, Pneumocystis
carinii, Trichomonas vaginalis, Histomonas meleagridis, Secementea, Trichuris
trichiura, Ascaris lumbricoides, Enterobius vermicularis, Ancylostoma duodenale,
Naegleria fowleri, Necator americanus, Nippostrongylus brasiliensis, Strongyloides
stercoralis, Wuchereria bancrofti, Dracunculus medinensis, blood flukes, liver flukes,
intestinal flukes, lung flukes, Schistosoma mansoni, Schistosoma haematobium,
Schistosoma japonicum, Fasciola hepatica, Fasciola gigantica, Heterophyes
heterophyes, and Paragonimus westermani.
90
9. The method as claimed in claims 1 to 4, wherein the
compound is
(Compound 19) or a pharmaceutically acceptable salt thereof.