[go: up one dir, main page]

US20230148436A2 - Novel small molecule immune agonists and immune targeting compounds and application thereof - Google Patents

Novel small molecule immune agonists and immune targeting compounds and application thereof Download PDF

Info

Publication number
US20230148436A2
US20230148436A2 US17/044,659 US201917044659A US2023148436A2 US 20230148436 A2 US20230148436 A2 US 20230148436A2 US 201917044659 A US201917044659 A US 201917044659A US 2023148436 A2 US2023148436 A2 US 2023148436A2
Authority
US
United States
Prior art keywords
szu
immune
inhibitors
group
pharmaceutically acceptable
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
Granted
Application number
US17/044,659
Other versions
US12281117B2 (en
US20210038605A1 (en
Inventor
Guangyi Jin
Zhulin Wang
Li Tang
Current Assignee (The listed assignees may be inaccurate. Google has not performed a legal analysis and makes no representation or warranty as to the accuracy of the list.)
Shenzhen University
Original Assignee
Shenzhen University
Priority date (The priority date is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the date listed.)
Filing date
Publication date
Application filed by Shenzhen University filed Critical Shenzhen University
Assigned to SHENZHEN UNIVERSITY reassignment SHENZHEN UNIVERSITY ASSIGNMENT OF ASSIGNORS INTEREST (SEE DOCUMENT FOR DETAILS). Assignors: JIN, GUANGYI, TANG, LI, WANG, Zhulin
Publication of US20210038605A1 publication Critical patent/US20210038605A1/en
Publication of US20230148436A2 publication Critical patent/US20230148436A2/en
Application granted granted Critical
Publication of US12281117B2 publication Critical patent/US12281117B2/en
Active legal-status Critical Current
Adjusted expiration legal-status Critical

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07D—HETEROCYCLIC COMPOUNDS
    • C07D473/00—Heterocyclic compounds containing purine ring systems
    • C07D473/02—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6
    • C07D473/18—Heterocyclic compounds containing purine ring systems with oxygen, sulphur, or nitrogen atoms directly attached in positions 2 and 6 one oxygen and one nitrogen atom, e.g. guanine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52—Purines, e.g. adenine
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K31/00—Medicinal preparations containing organic active ingredients
    • A61K31/33—Heterocyclic compounds
    • A61K31/395—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins
    • A61K31/495—Heterocyclic compounds having nitrogen as a ring hetero atom, e.g. guanethidine or rifamycins having six-membered rings with two or more nitrogen atoms as the only ring heteroatoms, e.g. piperazine or tetrazines
    • A61K31/505—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim
    • A61K31/519—Pyrimidines; Hydrogenated pyrimidines, e.g. trimethoprim ortho- or peri-condensed with heterocyclic rings
    • A61K31/52—Purines, e.g. adenine
    • A61K31/522—Purines, e.g. adenine having oxo groups directly attached to the heterocyclic ring, e.g. hypoxanthine, guanine, acyclovir
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K39/395—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum
    • A61K39/39533—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals
    • A61K39/3955—Antibodies; Immunoglobulins; Immune serum, e.g. antilymphocytic serum against materials from animals against proteinaceous materials, e.g. enzymes, hormones, lymphokines
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/545—Heterocyclic compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/549—Sugars, nucleosides, nucleotides or nucleic acids
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/54—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound
    • A61K47/55—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an organic compound the modifying agent being also a pharmacologically or therapeutically active agent, i.e. the entire conjugate being a codrug, i.e. a dimer, oligomer or polymer of pharmacologically or therapeutically active compounds
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/62—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being a protein, peptide or polyamino acid
    • A61K47/64—Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6801—Drug-antibody or immunoglobulin conjugates defined by the pharmacologically or therapeutically active agent
    • A61K47/6803—Drugs conjugated to an antibody or immunoglobulin, e.g. cisplatin-antibody conjugates
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K47/00—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient
    • A61K47/50—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates
    • A61K47/51—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent
    • A61K47/68—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment
    • A61K47/6835—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site
    • A61K47/6849—Medicinal preparations characterised by the non-active ingredients used, e.g. carriers or inert additives; Targeting or modifying agents chemically bound to the active ingredient the non-active ingredient being chemically bound to the active ingredient, e.g. polymer-drug conjugates the non-active ingredient being a modifying agent the modifying agent being an antibody, an immunoglobulin or a fragment thereof, e.g. an Fc-fragment the modifying agent being an antibody or an immunoglobulin bearing at least one antigen-binding site the antibody targeting a receptor, a cell surface antigen or a cell surface determinant
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/14—Antivirals for RNA viruses
    • A61P31/18—Antivirals for RNA viruses for HIV
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P31/00—Antiinfectives, i.e. antibiotics, antiseptics, chemotherapeutics
    • A61P31/12—Antivirals
    • A61P31/20—Antivirals for DNA viruses
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P35/00—Antineoplastic agents
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
    • A61P37/00—Drugs for immunological or allergic disorders
    • A61P37/02—Immunomodulators
    • A61P37/04—Immunostimulants
    • C—CHEMISTRY; METALLURGY
    • C07—ORGANIC CHEMISTRY
    • C07K—PEPTIDES
    • C07K16/00—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies
    • C07K16/18—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans
    • C07K16/28—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants
    • C07K16/2803—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily
    • C07K16/2827—Immunoglobulins [IGs], e.g. monoclonal or polyclonal antibodies against material from animals or humans against receptors, cell surface antigens or cell surface determinants against the immunoglobulin superfamily against B7 molecules, e.g. CD80, CD86
    • A—HUMAN NECESSITIES
    • A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
    • A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
    • A61K39/00—Medicinal preparations containing antigens or antibodies
    • A61K2039/505—Medicinal preparations containing antigens or antibodies comprising antibodies

Definitions

  • the present invention relates to a series of small-molecule immune agonists which are homologues, and to immune targeted compounds which are obtained by conjugating the agonists with targeted drugs and their applications, belonging to the field of an interdisciplinary science covering medicinal chemistry and immunology.
  • TLRs Toll-like receptors
  • ssRNAs single-stranded RNAs
  • TLR7 can be activated by artificially-synthesized small-molecule compounds to induce immunomodulation of the organism, and thus plays an important role in both anti-viral and anti-tumor therapies (Huju Chi, Chunman Li, Flora Sha Zhao, Li Zhang, Tzi Bun Ng, Guangyi Jin and Ou Sha. Anti-tumor Activity of Toll-Like Receptor 7 Agonists. Front Pharmacol. 2017 May 31; 8:304. doi:10.3389/fphar.2017.00304).
  • Tyrosine kinase inhibitors as the representatives of anti-tumor targeted therapeutic agents, can prolong the lives of patients; however, most patients will develop drug resistance within about one year or less.
  • TKIs Tyrosine kinase inhibitors
  • TLR7 immune agonists in earlier studies we had explored the combined administration of TKIs and TLR7 immune agonists in an attempt to achieve immune-targeted synergistic effects. Nevertheless, the laboratory results showed that the TKIs antagonized the immunostimulatory effects of TLR7 agonist on immunocytes and inhibited the production of immunostimulatory cytokines (Scientific Reports, 2016 Dec. 21; 6:39598.doi:10.1038/srep39598).
  • the objective of the present invention is on one hand to provide a series of small-molecule immune agonists having immunostimulatory effects and their applications, and on the other hand to provide immune targeted compounds obtained by conjugating said small-molecule immune agonists with targeted drugs and their applications.
  • the present invention relates to small-molecule immune agonists of Formula (I), or isomers or pharmaceutically acceptable salts thereof:
  • a 1 represents O or C
  • a 2 represents a nitro, amino, isocyanate, alkyleneamino, alkylene isocyanate, or a group selected from Formula (II) or (III):
  • R 1 and R 2 each independently represent (CH 2 ) p (NH) q , where p is 0, 1 or 2, and q is 0, 1 or 2;
  • R 3 represents O or S
  • R 5 represents a hydroxy, alkoxy, hydroxylamine, vinyl group or a group selected from:
  • R 6 represents alkyl or a group selected from:
  • the isomer is a geometric isomer or an enantiomer.
  • the present invention relates to small-molecule immune agonists of formula (I), or isomers or pharmaceutically acceptable salts thereof:
  • a 1 represents O or C
  • a 2 represents a nitro, amino, isocyanate, C 1 -C 6 alkylene amino, C 1 -C 6 alkylene isocyanate or a group selected from formula (II) or (III):
  • R 1 and R 2 each independently represent (CH 2 ) p (NH) q , where p is 0 or 1; and q is 0 or 1;
  • R 3 represents O or S
  • R 5 represents a hydroxyl, C 1 -C 6 alkoxy, hydroxylamine, vinyl group or a group selected from:
  • R 6 represents C 1 -C 6 alkyl group or a group selected from:
  • the present invention relates to small-molecule immune agonists of Formula (I), or isomers or pharmaceutically acceptable salts thereof:
  • a 1 represents O or C
  • a 2 represents a nitro, amino, isocyanate, methylene amino, methylene isocyanate or a group selected from Formula (II) or Formula (III):
  • R 1 and R 2 each independently represent (CH 2 ) p (NH) q , where p is 0 or 1, and q is 0 or 1;
  • R 3 represents O or S
  • R 5 represents a hydroxyl, methoxy, hydroxylamine, vinyl group or a group selected from:
  • R 6 represents a methyl group or a group selected from:
  • the present invention relates to small-molecule immune agonists of Formula (I), or isomers or pharmaceutically acceptable salts thereof, which are SZU-series compounds selected from the group consisting of (see Table 1 shown below for specific structural formulae): SZU-104, SZU-105, SZU-107, SZU-108, SZU-109, SZU-110, SZU-111, SZU-112, SZU-113, SZU-115, SZU-118, SZU-120, SZU-127, SZU-128, SZU-129, SZU-131, SZU-132, SZU-133, SZU-134, SZU-135, SZU-136, SZU-137, SZU-138, SZU-139, SZU-140, SZU-142, SZU-143, SZU-144, SZU-145, SZU-149, SZU-158, SZU-159, SZU-series
  • the present invention relates to immune targeted compounds or pharmaceutically acceptable salts thereof, which are formed by covalently conjugating the small-molecule immune agonists or an addition product formed from the immune agonists and mercapto-containing polypeptides such as glutathione, with targeted drugs or antibodies, and represented by the following general formula:
  • TLA represents small-molecule immune agonists or their addition products with mercapto-containing polypeptides such as glutathione
  • Tar represents targeted drugs or antibodies
  • n and m each independently represent a numerical value ranging from 1 to 5
  • L represents a linker, which is linear or branched alkyl or polyethylene glycol chain, and both ends of L are covalently linked to TLA and/or Tar directly or via additional groups.
  • the small-molecule immune agonists comprise the small-molecule immune agonists as described in the first aspect of the present invention and compounds selected from the group consisting of:
  • the small-molecule immune agonists are SZU-series compounds selected from the group consisting of (see Table 1 shown below for specific structural formulae): SZU-102, SZU-104, SZU-105, SZU-106, SZU-107, SZU-108, SZU-109, SZU-110, SZU-111, SZU-112, SZU-113, SZU-115, SZU-118, SZU-120, SZU-127, SZU-128, SZU-129, SZU-131, SZU-132, SZU-133, SZU-134, SZU-135, SZU-136, SZU-137, SZU-138, SZU-139, SZU-140, SZU-142, SZU-143, SZU-144, SZU-145, SZU-149, SZU-158, SZU-159, SZU-160, SZU-161, SZU-series
  • the TLA represents compounds selected from the group consisting of (see Table 3 shown below for specific structural formulae): SZU-103-GSH, SZU-114-GSH, SZU-117-GSH, SZU-122-GSH, SZU-127-GSH, SZU-133-GSH, SZU-138-GSH and SZU-144-GSH.
  • the Tar represents at least one compound selected from the group consisting of: Dasatinib, Imatinib, Saracatinib, Ponatinib, Nilotinib, Danusertib, AT9283Degrasyn, Bafetinib, kw-2449, nyp-bhg712, dcc-2036, GZD824, GNF-2, PD173955, GNF-5, Bosutinib, Gefitinib, Erlotinib, Sunitinib, Ruxolitinib, Tofacitinib, Lapatinib, Vandetanib, Sorafenib, Sunitinib, Axitinib, Nintedanib, Regorafenib, Pazopanib, Lenvatinib, Crizotinib, Ceritinib, Cabozantinib, DWF, Afatinib, Ibrutinib, Nirap
  • the Tar represents at least one of the compounds selected from the group consisting of: TKI, EGFR inhibitors, KRAS inhibitors, BRAF inhibitors, BTK inhibitors, BRD4 inhibitors, PD-L1 inhibitors, PD-1 inhibitors, OX40 agonists, PARP inhibitors and CDK4/6 inhibitors.
  • L is
  • the additional groups include amides, alkoxy bonds, alkylthio bonds, substituted amine groups, quaternary amine salts, heterocycles formed by Click reaction, amino acid groups that are dissociable by proteases, ureido or thiourea groups.
  • the immune targeted compounds or pharmaceutically acceptable salts thereof are SZU-series compounds selected from the group consisting of (see Table 4 shown below for specific structural formulae): SZU-116, SZU-119, SZU-124, SZU-125, SZU-146, SZU-147, SZU-168, SZU-169, SZU-174, SZU-175, SZU-176, SZU-177, SZU-178, SZU-179, SZU-180, SZU-181, SZU-158-PD-L1, SZU-158-OX40, SZU-158-PD-1 or SZU-136-miRNA21, or pharmaceutically acceptable salts thereof.
  • the present invention relates to use of the small-molecule immune agonists of formula (I) or isomers or pharmaceutically acceptable salts thereof according to the first aspect of the present invention, compounds SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130 (see Table 2 shown below for specific structural formulae) and the immune targeted compounds or pharmaceutically acceptable salts thereof according to the second aspect of the present invention, in the manufacture of a medicament that participates in immunomodulation.
  • the present invention relates to use of the small-molecule immune agonists of formula (I) or isomers or pharmaceutically acceptable salts thereof according to the first aspect of the present invention, compounds SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130 and the immune targeted compounds or pharmaceutically acceptable salts thereof according to the second aspect of the present invention, in the manufacture of a medicament as an anti-tumor or anti-viral drug or for eliminating target proteins.
  • the present invention further relates to use of a compound SZU-136-miRNA21 and analogues thereof in the manufacture of a medicament for immunologically targeting RNA, wherein the compound SZU-136-miRNA21 and analogues thereof are useful as small molecular RNAs for immune targeting; and the medicament may be present in various dosage forms, including solid preparations, liquid preparations, and conjugates formed from the medicament with various vehicles or crystalline hydrates of the medicament.
  • the present invention further relates to use of compounds SZU-179, SZU-180 and SZU-181 in the manufacture of a medicament as anti-AIDS and anti-hepatitis B agents, wherein the compounds SZU-179, SZU-180 and SZU-181 are useful as antiviral immune targeted compound.
  • the present invention further relates to use of compounds SZU-161 and SZU-162 for determining the conjugation degree of the small-molecule immune agonists conjugated with protein drugs and cells, wherein the compounds SZU-161 and SZU-162 are useful as small-molecule immune agonists with specific ultraviolet absorption.
  • the present invention further relates to use of compounds SZU-116 and SZU-124 in observing and studying the cellular absorption and distribution in vivo of proteins or targeted drugs, wherein compounds SZU-116 and SZU-124 are conjugated with proteins or targeted drugs, and are useful as compounds with optical tracking properties of chromogenic indicators for small-molecule immune agonists. After having been conjugated with proteins or targeted drugs, they can show the cellular absorption and distribution in vivo of proteins or targeted drugs in real time.
  • the present invention further relates to use of compounds SZU-114, SZU-117, SZU-122, SZU-127, SZU-133, SZU-138, and SZU-143 in the manufacture of a medicament with dual immune targeted functions, wherein the compounds SZU-114, SZU-117, SZU-122, SZU-127, SZU-133, SZU-138, and SZU-143 are conjugated with targeted drugs or antibodies.
  • the small-molecule immune agonists of the present invention per se have immunostimulatory effects. Furthermore, it is well-known to the skilled persons that the combined application of TKI and TLR7 immune agonist will cause TKIs to antagonize the activation of TLR7 agonists on immunocytes, thereby inhibiting the production of immunostimulatory cytokines.
  • TKIs as the targeted drugs
  • TLR7 small-molecule immune agonists the inventors have surprisingly found that such covalently conjugated compounds not only maintain or improve the immunostimulatory effects of the small-molecule immune agonists, but also enhance the anti-tumor effects of the targeted drugs.
  • the immune targeted compounds obtained by the method of the present invention are of great significance for the long-term and high-effective treatment of tumors.
  • novel small-molecule immune agonists that have immunostimulatory effects and can be conjugated with targeted drugs to form immune targeted compounds with targeting action as described above are shown in Table 1 below:
  • Some small-molecule immune agonists listed in Tables 1 and 2 can form addition products with mercapto-containing polypeptides such as glutathione (GSH), which can target glutathione transferase, result in improved therapeutic efficacy on diseases by acting synergistically with chemotherapy drugs, irreversible TKI targeted drugs and other irreversible targeted drugs, and eliminate drug resistance, while activating immunological effects.
  • GSH glutathione
  • SZU-117-GSH formed by reacting SZU-117 with glutathione via a synthesis process as shown in the following scheme, is illustrated as an example.
  • Examples of the addition products formed from the small-molecule immune agonists and GSH are shown in Table 3:
  • mercapto-containing compounds include, for example, 2-methylamino ethanethiol and
  • the small-molecule immune agonists listed in Tables 1 and 2 and the compounds formed from some small-molecule immune agonists and GSH listed in Table 3 can be conjugated with targeted drugs to give bi-functional immune targeted compounds with immune targeting effects.
  • the immune targeted compounds obtained are shown in Table 4 below:
  • Small-molecule immune agonists are conjugated with targeted drugs to form immune targeted compounds via synthesis schemes shown below:
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme I:
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme II:
  • the same process as described above may also be applied to conjugating of SZU-144 or SZU-133 with targeted drugs to form immune targeted compounds.
  • the linker may also be a simple linear alkane or other linkers replaced as desired.
  • Carboxyl-containing small-molecule immune agonists such as SZU-138 can be conjugated with antibodies through addition reactions and conjugation reactions to form soluble, functional protein polypeptide conjugates, which can be formed into salts owing to the presence of the dimethylamino group, so that they can provide ionized positive charges, and thus are advantageous for increasing the therapeutic efficacy and water solubility, thereby solving the solubility problem of the conjugated products.
  • carboxyl-containing small-molecule immune agonists as described above can also be conjugated with two different targeted drugs to form dual-targeted immune compounds.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme III:
  • Products formed from small-molecule immune agonists and glutathione can also be conjugated with two different targeted drugs to enhance the therapeutic efficacy.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme IV:
  • the targeted drugs can either be various TKI agents and tumor targeted drugs, or antibodies such as CD3 antibody, PD-1 antibody, OX40 antibody, CD19 antibody, CD20 antibody, HER2 antibody, MUC1 antibody and antibodies against various tumor proteins and pathogens, wherein the linker may be alkane or alkoxy chains, and can be adjusted depending on the requirements for optimizing dissolution and metabolism.
  • Small-molecule immune agonists (using SZU-166 as an example, which is a unique isocyanate small-molecule immune agonist with the merit of mild quantitative conjugate addition with amino compounds) may be conjugated with targeted drugs via Synthesis Scheme V:
  • Compounds SZU-115 and SZU-160 are novel small-molecule immune agonists with Click reactive groups that mainly have two functions including: 1) after having been conjugated with macromolecules and biologically active substances, being useful in optical tracking of the conjugated product by linking to a chromogenic indicator via Click reaction; for example, when they have conjugated with proteins or targeted drugs, the cellular absorption and distribution in vivo of the proteins and targeted drugs are observed.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme VI:
  • SZU-115 can also be simultaneously linked to antibodies and targeted drugs, thereby leading to reduced off-target effects of the antibodies, enhanced efficacy of the targeted drugs and activated antitumor immunity:
  • Compound SZU-139 in the present invention is prepared by Click reaction of SZU-136 with ribose:
  • miRNA21 plays an important role in generation and development of tumor (Cell Death Dis. 2018 Feb. 13; 9(2):219. doi: 10.1038/s41419-017-0243-9; Proc Natl Acad Sci S U.S.A. 2015 Jun. 30; 112(26): E3355-64. doi: 10.1073/pnas.1504630112; Cancer Lett. 2017 May 1; 393: 86-93.doi:10.1016/j.canlet.2017.02.019; https://en.wikipedia.org/wiki/MIRN21).
  • the application of small-molecule immune agonists conjugated with pathogenic miRNAs can guide the immune system to block the occurrence of diseases at source, the changes of related proteins and the onset of related diseases.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme VII:
  • the alkyne immune agonist can be replaced with SZU-115.
  • the carboxyl group of SZU-115 can be conjugated with Argonaute to form a conjugate, which interrupts the formation of a conjugate of miRNA and Argonaute, and is directed into the endosome of immunocytes under the action of TLR7 agonist, thus resulting in immunological elimination and memory:
  • the inventors have found a series of small-molecule immune agonists, which can be conjugated with targeted drugs to form novel immune targeted compounds with immunostimulatory function, thereby resulting in increased in vitro and in vivo immunostimulatory effects of the targeted drugs and improved efficacy against tumors and other diseases, conferring dual functions to the targeted drugs (bi-functional targeted drugs).
  • Such an enhanced effect is resulted from the synergistic action of two functions: inclusion of antitumor immune factors (such as IFN- ⁇ ) and inhibition on the pathogenic targeted sites.
  • the present invention is directed to novel high-effective targeted drugs with a combination of the two functions.
  • the immune targeted compounds according to the present invention act appropriately by eliminating pathogenic targets (proteins) via immune pathway as shown in FIG. 70 .
  • Immune targeted compounds formed by conjugating small-molecule immune agonists with targeted drugs are guided by the targeted drugs to the pathogenic proteins, thereby resulting in the death of the targeted cells and release of the pathogenic proteins. Further, they are guided by the immune agonists to antigen presenting cells (such as DC), in which the pathogenic proteins are cleaved, leading to degradation of these proteins.
  • the immune targeted compounds formed by conjugating small-molecule immune agonists with targeted drugs (such as TKIs) in the present invention can overcome drug resistance and improve therapeutic efficacy.
  • Lysine-derived compounds SZU-104, SZU-105, SZU-112 and SZU-113 that are useful as acidic small-molecule immune agonists are amino acid derivatives having the activity of immune agonists, and can achieve the purpose of site-specific conjugation by inserting polypeptides (or antibodies) at specific sites as desired in the preparation of immunogenic polypeptides:
  • SZU-161 and SZU-162 as small-molecule immune agonists suitable for conjugating with protein drugs, have a specific ultraviolet absorption peak (342 nM), and can be used typically for measuring the conjugation degree of small-molecule immune agonists conjugated with proteins drugs:
  • SZU-161 was measured for the intensity of optical absorption at 342 nM under different concentrations; standard curves of concentration and absorption were plotted to obtain the concentration-absorption relationship formula; and 2) after conjugation with protein, the conjugated protein was measured for the absorption at 342 nM under certain concentration; the resulting value was placed into the relationship formula to readily obtain the relative concentration of SZU-161 in the conjugate (i.e., conjugation degree).
  • SZU-162 may be applied to achieve the same effect.
  • immunocytokines refer to cytokines (including IFN- ⁇ , IL-12, IL-6, and TNF- ⁇ ), and their concentration levels can be used to indicate the levels of immunocyte responsiveness stimulated by the compound.
  • FIG. 1 shows the stimulatory effects of SZU-GSH-series compounds on immune cytokine (IL-6).
  • FIG. 2 shows the distribution of fluorescence-labeled SZU-116 in cells.
  • FIG. 3 shows the stimulatory effects of SZU-102, 103, 106, 107, 108, 110, 111, 114, 115, 116, 117, and 118 on immune cytokine (IFN- ⁇ ), wherein SZU-101 and R848, as the standard control, are internationally-accepted, standard small-molecule immune agonists with immunostimulatory function (the same below).
  • IFN- ⁇ immune cytokine
  • FIG. 4 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (IFN- ⁇ ), wherein “con” is PBS blank control (the same below).
  • FIG. 5 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (IL-12).
  • FIG. 6 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (TNF- ⁇ ).
  • FIG. 7 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (IL-6).
  • FIG. 8 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokine (IFN- ⁇ ).
  • FIG. 9 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokine (IL-12).
  • FIG. 10 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokine (TNF- ⁇ ).
  • FIG. 11 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokines (IL-6).
  • FIG. 12 shows the stimulatory effects of SZU-125 on several immune cytokines.
  • FIG. 13 shows the stimulatory effects of SZU-105 on immune cytokine (IFN- ⁇ ).
  • FIG. 14 shows the stimulatory effects of SZU-105 on immune cytokine (IL-12).
  • FIG. 15 shows the stimulatory effects of SZU-113 on immune cytokine (IFN- ⁇ ).
  • FIG. 16 shows the stimulatory effects of SZU-113 on immune cytokine (IL-12).
  • FIG. 17 shows the stimulatory effects of SZU-119 on immune cytokine (IL-12).
  • FIG. 18 shows the stimulatory effects of SZU-119 on immune cytokine (IL-6).
  • FIG. 19 shows the stimulatory effects of SZU-120 on immune cytokine (IFN- ⁇ ).
  • FIG. 20 shows the stimulatory effects of SZU-120 on immune cytokine (IL-6).
  • FIGS. 21 - 54 show the stimulatory effects of the small-molecule immune agonists SZU-122 and 127-142 according to the present invention on immune cytokines, respectively.
  • FIG. 55 shows the stimulatory effects of SZU-102, 119, 125, 146, 147, 169, 174, 175, 168, 176, 177, and 179 on immunocytes to produce IFN- ⁇ .
  • FIG. 56 shows the stimulatory effects of SZU-102, 119, 125, 146, 147, 169, 174, 175, 168, 176, 177, and 179 on immunocytes to produce IL-12.
  • FIG. 57 shows the anti-tumor (CT26) activity of novel immune targeted compounds in vivo: Veh (untreated control), A (SZU-177), B (SZU-175), C (SZU-174), D (SZU-147), and E (SZU-176).
  • CT26 anti-tumor
  • FIG. 58 shows the anti-tumor (B16) activity of novel immune targeted compounds in vivo: Veh (untreated control), G (PD-L1 antibody), H (SZU-178), I (SZU-158-PD-L1), J (SZU-158-OX40), K (PD-1 antibody), and L (SZU-158-PD-1).
  • FIG. 59 shows the inhibition of JQ1 on the activity of SZU-101 as a TLR7 agonist; wherein SZU-119 retains the activity of TLR7 agonist for stimulating immune cytokine.
  • FIG. 60 shows the inhibition of JQ1 on the activity of SZU-101 as a TLR7 agonist, wherein SZU-119 retains the activity of TLR7 agonist for stimulating immune cytokine.
  • FIG. 61 shows that SZU-119 retains the inhibitory activity of JQ1 against CD274 (PD-L1) on B16 cells, wherein “control” represents blank control.
  • FIG. 62 shows the stimulatory effects of SZU-163 and SZU-166 on immunocytes to produce IFN- ⁇ .
  • FIG. 63 shows the stimulatory effects of SZU-180 and SZU-136-miRNA21 on immunocytes to produce IFN- ⁇ .
  • FIG. 64 shows the anti-tumor (4T1) survival rates of PD-1 antibody, PD-L1 antibody, OX40 antibody and their SZU-158 conjugates and SZU-178.
  • FIG. 65 shows the stimulatory effects of PD-1 antibody, PD-L1 antibody, OX40 antibody, and their SZU-158 conjugates and SZU-178 on immunocytes to produce IFN- ⁇ .
  • FIG. 66 shows the stimulatory effects of SZU-181 on immunocytes to produce IFN- ⁇ .
  • FIG. 67 shows the stimulatory effects of SZU-106, SZU-158, and SZU-160 on immunocytes to produce IFN- ⁇ , wherein “control” represents PBS blank control.
  • FIG. 68 shows the stimulatory effects of SZU-106, SZU-158, and SZU-160 on immunocytes to produce IL-6, where “control” represents blank control.
  • FIG. 69 is a schematic diagram showing the functioning mechanism of TLR7 small-molecule immune agonist conjugated with self-miRNAs in human bodies for immune-targeted interference in the binding of agonist to protein, thereby eliminating protein-related pathogenicity.
  • FIG. 70 is a schematic diagram showing the functioning mechanism of the immune targeted compound of the present invention for eliminating pathogenic targets (proteins) via immune pathway.
  • the SZU-030T obtained in the above step was dissolved in 10 mL of anhydrous tetrahydrofuran (THF). 484 mg of triphenylphosphine was added under an ice bath. The mixture was transferred to room temperature and continued to react under stirring overnight. The progress of the reaction was monitored by LC-MS. After the reaction was completed, 20 mL of water was added. The mixture was stirred for another 30 minutes, then filtered off with suction under reduced pressure. The residue was washed with water twice and dried to give 625 mg of SZU-031 T with a yield of 79% over the two steps.
  • THF anhydrous tetrahydrofuran
  • SZU-124 was prepared by the same process as SZU-116, except that SZU-017T was replaced with FITC, and a grass green semi-solid was obtained. The yield over the two steps was 31%.
  • ESI-MS: m/z 1199.5 [M+H] + .
  • SZU-127 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with retinoic acid, and a yellow solid was obtained.
  • ESI-MS: m/z 627.4[M+H] + .
  • SZU-134 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with glutaric anhydride; and a white solid was obtained.
  • ESI-MS: m/z 459.1[M+H] + .
  • SZU-138 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with itaconic anhydride; and a white solid was obtained.
  • ESI-MS: m/z 457.1[M+H] + .
  • SZU-140 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with acetic anhydride; and a white solid was obtained.
  • ESI-MS: m/z 387.1[M+H] + .
  • SZU-114-GSH was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-114; and a white solid was obtained.
  • ESI-MS: m/z 763.3[M+H] + .
  • SZU-117-GSH was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-117; and a white solid was obtained.
  • ESI-MS: m/z 874.3[M+H] + .
  • SZU-158 was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-138; glutathione was replaced with 2-(dimethylamino)ethane-1-thiol; and white solid was obtained.
  • SZU-159 can be prepared by the same process, except that SZU-138 was replaced with SZU-144, which was reacted with 2-(dimethylamino)ethane-1-thiol at room temperature for 24 hours to give SZU-159.
  • SZU-161 was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-138; glutathione was replaced with SZU-033T; and a white solid was obtained.
  • ESI-MS: m/z 708.4[M+H] + .
  • SZU-162 was prepared by the same process as SZU-161.
  • ESI-MS: m/z 695.81[M+H] + .
  • SZU-160 was prepared by the same process as SZU-115, except that SZU-013T was replaced with SZU-032T; and a white solid was obtained.
  • ESI-MS: m/z 599.3[M+H] + .
  • Lithium hydroxide and SZU-101 (in 1:1 ratio) were added to water. The resulting solution was stirred for 2-3 hours, and then lyophilized to remove the solvent to obtain a pure product.
  • Bromine was added dropwise slowly to the acetic acid solution of SZU-151. After stirring overnight, the reaction solution was neutralized with NaHCO 3 . Small amount of ethanol was added into the obtained viscous compound. Subsequently, sodium thiosulfate solution was added to remove the remaining bromine. The reaction solution was filtered off with suction and dried to give a solid, and then 10% NaOH solution was added. The resulting solution was further filtered off with suction and dried under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a bromide.
  • SZU-134 was dissolved in DMSO. Equimoles of NHS and EDCI were added. After stirring for 10 minutes, an equimole of Direct violet N (OX40 agonist) was added. The reactants were reacted in a closed vessel at room temperature for 3 days. 20-fold amount of water was added. Solid was precipitated out and filtered, and then 2N NaOH was added. The mixture was stirred at 10° C. for 30 minutes and filtered. The filtrate was acidified with 2N hydrochloric acid to have a pH of 1 and gave a precipitated solid product SZU-169 with a yield of 52%; MS (ESI): m/z: [M+2Na] 1611.64.
  • the reactant a was dissolved in small amount of DMF. One-fold equivalent amount of sodium azide was added. The mixture was reacted under stirring at room temperature for 24 hours. The reaction mixture was added with 10-fold volume of water and extracted with ethyl acetate. The extracted phase was washed with water three times. The solvent was distilled off under reduced pressure to obtain product b with a yield of 90%.
  • novel immune-targeted compounds formed by conjugating SZU-158 with an antibody owing to the presence of dimethylamino group, can form a salt and result in an ionized positive charge, which is advantageous for improving the therapeutic efficacy and water solubility.
  • Vemu-1 a precursor of Vemurafenib, is purchased from WuXi AppTec.
  • Vemu-1 was dissolved in DMF. 2-fold moles of triethylamine were added. The mixture was cooled to 0° C. and then added with 1-fold mole of vinylsulfonyl chloride. The resulting mixture was warmed slowly to room temperature, stirred for 8 hours, and then added with 10-fold volume of ice-water. The mixture was stirred thoroughly for 1 hour and adjusted with saturated Na 2 CO 3 to have a pH of 8. The precipitated solid was filtered, washed with water twice, and dried to give Vemu-2. 1 g of Vemu-2 was dissolved in 30 mL of DMF. 0.33 g of mercaptoethylamine was added. The mixture was stirred at room temperature for 12 hours.
  • SZU-160 was dissolved in 0.5 mL of DMSO. Equimoles of NHS and EDCI were added. The mixture was left under stirring at room temperature for 4 hours. 20 mg of PD-L1 antibody (purchased from Bioxcell, 10F.9G2) in 0.5 mL of PBS was added. The mixture was stirred in a closed vessel at 15° C. overnight. The mixed reaction solution was filtered with a filter membrane to obtain a solution of SZU-160-PD-L1 antibody conjugate product. 2.5 mg of Erlotinib, 2 mg of sodium ascorbate, and 2 mg of copper sulfate were added. The mixture was reacted under stirring at room temperature for 12 hours.
  • the reaction solution was filtered with filter membrane and lyophilized to give 12 mg of a white solid SZU-178 (i.e., SZU-160-Erlotinib-PD-L1 conjugate formed by conjugating SZU-178 with Erlotinib and PD-L1 antibody).
  • SZU-115-PD-L1-targeted drug i.e., the conjugate formed by conjugating SZU-115 with PD-L1 antibody and targeted drug
  • the PD-L1 antibody was conjugated by the same process as in the preparation of SZU-178, and Click reaction was conducted by the same process as in the preparation of SZU-179, to obtain SZU-180.
  • the resulting product can be used for the treatment of AIDS.
  • BALB/c or C57BL/6 mouse spleens were collected, ground in a small dish containing Mouse Lymphocyte Separation Medium, and filtered through a filter membrane (with a pore size of 0.22 ⁇ m) to obtain a cell suspension, which was treated by centrifugation to obtain lymphocytes.
  • the isolated lymphocytes were placed in a 15 mL centrifuge tube, into which 10 mL of 1640 complete medium was added. The mixture was mixed uniformly and centrifuged at 250 g for 10 minutes. The supernatant was discarded.
  • the spleen lymphocytes seeded into the 24-well plates were stimulated with different concentrations of the SZU-series small-molecule immune agonists and novel immune targeted compounds.
  • the 24-well plate was transferred into an incubator. After 24 hours of stimulation, the supernatant was gently aspirated, i.e. samples to be tested by ELISA (Enzyme-Linked Immunosorbent Assay). The concentrations and concentration gradients were dependent on individual requirements of each tested sample.
  • Coating 10 ⁇ coating buffer was diluted to 1 ⁇ and then used to dilute the capture antibody.
  • the diluted capture antibody was added into a 96-well microtiter plate, 100 ⁇ l per well.
  • the plate was sealed with plastic wrap and placed at 37° C. for 2 to 4 hours or at 4° C. in refrigerator overnight. The liquid contents were aspirated out of each well of the plate. Then the plate was washed with washing buffer (PBST:PBS solution with 5% Tween-20) three times and dried by patting.
  • washing buffer PBST:PBS solution with 5% Tween-20
  • Blocking diluted blocking solution was added, 200 ⁇ l per well; the plate was sealed with plastic wrap and blocked at room temperature for 1 hour. Then the plate was washed with PBST three times and dried by patting.
  • Loading samples 12 standard samples were arranged, wells in duplicate for each concentration of a standard sample, with the highest concentration of the standard sample being 2,000 pg/mL. The samples were diluted with dilution buffer to the desired concentration. The standard samples have the following successive concentrations: 2,000 pg/mL, 1,000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.5 pg/mL, 15.75 pg/mL, 7.875 pg/mL, 3.9375 pg/mL, 1.96875 pg/mL, and 0 pg/mL.
  • the supernatant samples obtained by stimulating spleen lymphocytes with different concentrations of drugs were added into the corresponding wells, wells in triplicate per sample. Blank control wells and negative control wells were also arranged and labeled accordingly.
  • the 96-well microtiter plate loaded with the standard samples and tested samples was sealed with plastic wrap and incubated at room temperature for 2 hours or at 4° C. in refrigerator overnight. The liquid contents were aspirated out of each well. The plate was washed with PBST three times and dried by patting.
  • the testing antibody was diluted with dilution buffer to the desired concentration, and added to the plate at 100 ⁇ l per well, and incubated at room temperature for 1 hour. The liquid contents were aspirated out of each well of the plate. Then the plate was washed with PBST five times and dried by patting.
  • Enzyme-labeled antibodies Avidin-HRP was diluted with dilution buffer to the desired concentration, and added to the plate at 100 ⁇ l per well, and then incubated at room temperature for 30 minutes. The liquid contents were aspirated out of each well of the plate. Then the plate was washed with PBST five times and dried by patting.
  • TMB 3,3′,5,5′-tetramethylbenzidine
  • Termination after the completion of color development, 50p of stop solution was added to each well, wherein the stop solution was 1 mol/L H 2 SO 4 .
  • mice of 6-8 weeks old were weighed and anesthetized with pentobarbital sodium (50 mg/kg i.p). Hair was removed from the tumor implantation site (on the back). The mice were immobilized with sticky tapes.
  • Surgical sites were sterilized by using cotton swabs dipped into 75% alcohol. Small incisions of about 1 cm at the tumor implantation sites were made and injected with 100 ⁇ l of cancer cell suspension (5 ⁇ 104 CT26 cells, B16 cells or 4T1 cells). The incisions were sealed with surgical thread and sterilized. After the operations, 50,000 units of ampicillin/100 ⁇ L/mouse were injected intraperitoneally for 3 consecutive days.
  • mice were administered with PBS blank control, the immune agonists or novel immune targeted compounds, respectively.
  • the dosages were determined according to the actual needs (following the principle of effective and safe dosages).
  • the respective substance was dissolved in an appropriate solvent, and the volume of each administration was 100 ⁇ L; the mice in each group were administered via intraperitoneal injection.
  • Each group was administered on day 7, 15, 22, and 29 after tumor implantation; and the tumor sizes were measured regularly.
  • the mice were euthanized when the tumor size reached 1500 mm 3 or had a weight higher than 15% of their body weights.
  • the results of tumor suppression and survival rates were shown in FIGS. 57 , 58 and 64 . It can be seen that the groups of mice treated with the novel immune targeted compounds according to the present invention have significantly higher anti-tumor effect.
  • Methods for testing the biological activities of the novel small-molecule immune agonists and novel immune targeted compounds of the present invention are not limited to the above test methods, and other known or accepted activity testing methods may also be used.
  • the schematic diagrams of activities in the present invention are not limited to the illustrated schematic diagrams, while these schematic diagrams can inspire those skilled in the art to achieve the application effects of the novel small-molecule immune agonists and novel immune targeted compounds according to the present invention.

Landscapes

  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Veterinary Medicine (AREA)
  • Public Health (AREA)
  • Pharmacology & Pharmacy (AREA)
  • Animal Behavior & Ethology (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Organic Chemistry (AREA)
  • Immunology (AREA)
  • Epidemiology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • General Chemical & Material Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Virology (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Oncology (AREA)
  • Communicable Diseases (AREA)
  • Biochemistry (AREA)
  • AIDS & HIV (AREA)
  • Tropical Medicine & Parasitology (AREA)
  • Biotechnology (AREA)
  • Microbiology (AREA)
  • Mycology (AREA)
  • Cell Biology (AREA)
  • Endocrinology (AREA)
  • Biophysics (AREA)
  • Genetics & Genomics (AREA)
  • Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
  • Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
  • Acyclic And Carbocyclic Compounds In Medicinal Compositions (AREA)
  • Medicinal Preparation (AREA)

Abstract

Provided is a series of homologous small molecule immune agonists and novel bifunctional immune targeting compounds having targeting and immune activation functions, which are obtained by coupling the small molecule immune agonists to targeting drugs. The resulting immune targeting compounds are beneficial for enhancing immune activation effects, and anti-tumor and other disease fighting effects of the targeting drug. The enhanced effect is produced from a synergy of immunological anti-tumor factors (such as IFN-γ) and inhibition at pathogenic targeting sites.

Description

    CROSS-REFERENCE TO RELATED APPLICATIONS
  • This application is a U.S. National Phase Application of International Application No. PCT/CN2019/080945, filed Apr. 2, 2019, which claims priority to Chinese Application No. 201810289351.7, filed Apr. 3, 2018, and Chinese Application No. 201810331582.X, filed Apr. 13, 2018, each of which are hereby incorporated by reference in their entirety.
    • FIELD OF THE INVENTION
  • The present invention relates to a series of small-molecule immune agonists which are homologues, and to immune targeted compounds which are obtained by conjugating the agonists with targeted drugs and their applications, belonging to the field of an interdisciplinary science covering medicinal chemistry and immunology.
    • BACKGROUND OF THE INVENTION
  • Toll-like receptors (TLRs) present an organism's first line of defense in the defense system (Kaisho, T. &Akira, S. Toll-like receptor function and signaling. J. Allergy Clin. Immunol. 117, 979-987 (2006); Miyake, K. Innate immune sensing of pathogens and danger signals by cell surface Toll-like receptors. Semin. Immunol. 19, 3-10 (2007)). In particular, toll-like receptor 7 (TLR7), which is activated by single-stranded RNAs (ssRNAs) as its natural ligand, plays a crucial role in the defense against invasions of ssRNA viruses (Blasius, A. L. & Beutler, B. Intracellular toll-like receptors. Immunity 32, 305-315 (2010)). Furthermore, TLR7 can be activated by artificially-synthesized small-molecule compounds to induce immunomodulation of the organism, and thus plays an important role in both anti-viral and anti-tumor therapies (Huju Chi, Chunman Li, Flora Sha Zhao, Li Zhang, Tzi Bun Ng, Guangyi Jin and Ou Sha. Anti-tumor Activity of Toll-Like Receptor 7 Agonists. Front Pharmacol. 2017 May 31; 8:304. doi:10.3389/fphar.2017.00304).
  • Tyrosine kinase inhibitors (TKIs), as the representatives of anti-tumor targeted therapeutic agents, can prolong the lives of patients; however, most patients will develop drug resistance within about one year or less. In order to address the drug resistance of TKIs, in earlier studies we had explored the combined administration of TKIs and TLR7 immune agonists in an attempt to achieve immune-targeted synergistic effects. Nevertheless, the laboratory results showed that the TKIs antagonized the immunostimulatory effects of TLR7 agonist on immunocytes and inhibited the production of immunostimulatory cytokines (Scientific Reports, 2016 Dec. 21; 6:39598.doi:10.1038/srep39598).
    • SUMMARY OF THE INVENTION
  • The objective of the present invention is on one hand to provide a series of small-molecule immune agonists having immunostimulatory effects and their applications, and on the other hand to provide immune targeted compounds obtained by conjugating said small-molecule immune agonists with targeted drugs and their applications.
  • In the first aspect, the present invention relates to small-molecule immune agonists of Formula (I), or isomers or pharmaceutically acceptable salts thereof:
  • Figure US20230148436A2-20230511-C00001
  • wherein
  • A1 represents O or C;
  • A2 represents a nitro, amino, isocyanate, alkyleneamino, alkylene isocyanate, or a group selected from Formula (II) or (III):
  • Figure US20230148436A2-20230511-C00002
  • wherein
  • R1 and R2 each independently represent (CH2)p(NH)q, where p is 0, 1 or 2, and q is 0, 1 or 2;
  • R3 represents O or S;
  • R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0, 1 or 2; z is 0, 1 or 2; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0, 1 or 2, wherein when s≠0 and t=0, the carbon atom adjacent to the carbonyl group in (CH2)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
  • Figure US20230148436A2-20230511-C00003
  • or, or forming a double bond with another carbon atom;
  • R5 represents a hydroxy, alkoxy, hydroxylamine, vinyl group or a group selected from:
  • Figure US20230148436A2-20230511-C00004
  • R6 represents alkyl or a group selected from:
  • Figure US20230148436A2-20230511-C00005
  • where * denotes the site linked to the carbon atom;
  • with the proviso that the following compounds are excluded:
  • Figure US20230148436A2-20230511-C00006
    Figure US20230148436A2-20230511-C00007
  • Preferably, the isomer is a geometric isomer or an enantiomer.
  • In a preferred embodiment, the present invention relates to small-molecule immune agonists of formula (I), or isomers or pharmaceutically acceptable salts thereof:
  • Figure US20230148436A2-20230511-C00008
  • wherein
  • A1 represents O or C;
  • A2 represents a nitro, amino, isocyanate, C1-C6 alkylene amino, C1-C6 alkylene isocyanate or a group selected from formula (II) or (III):
  • Figure US20230148436A2-20230511-C00009
  • wherein
  • R1 and R2 each independently represent (CH2)p(NH)q, where p is 0 or 1; and q is 0 or 1;
  • R3 represents O or S;
  • R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0 or 1; z is 0 or 1; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0 or 1, wherein when s≠0 and t=0, the carbon atom adjacent to the carbonyl group in (CH2)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
  • Figure US20230148436A2-20230511-C00010
  • or forming a double bond with another carbon atom;
  • R5 represents a hydroxyl, C1-C6 alkoxy, hydroxylamine, vinyl group or a group selected from:
  • Figure US20230148436A2-20230511-C00011
  • R6 represents C1-C6 alkyl group or a group selected from:
  • Figure US20230148436A2-20230511-C00012
  • wherein * denotes the site linked to the carbon atom;
  • with the proviso that the following compounds are excluded:
  • Figure US20230148436A2-20230511-C00013
    Figure US20230148436A2-20230511-C00014
  • In a further preferred embodiment, the present invention relates to small-molecule immune agonists of Formula (I), or isomers or pharmaceutically acceptable salts thereof:
  • Figure US20230148436A2-20230511-C00015
  • wherein,
  • A1 represents O or C;
  • A2 represents a nitro, amino, isocyanate, methylene amino, methylene isocyanate or a group selected from Formula (II) or Formula (III):
  • Figure US20230148436A2-20230511-C00016
  • wherein,
  • R1 and R2 each independently represent (CH2)p(NH)q, where p is 0 or 1, and q is 0 or 1;
  • R3 represents O or S;
  • R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0 or 1; z is 0 or 1; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0 or 1, wherein when s≠0 and t=0, the carbon atom adjacent to the carbonyl group in (CH2)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
  • Figure US20230148436A2-20230511-C00017
  • or forming a double bond with another carbon atom;
  • R5 represents a hydroxyl, methoxy, hydroxylamine, vinyl group or a group selected from:
  • Figure US20230148436A2-20230511-C00018
  • R6 represents a methyl group or a group selected from:
  • Figure US20230148436A2-20230511-C00019
  • wherein * denotes the site linked to the carbon atom;
  • with the proviso that the following compounds are excluded:
  • Figure US20230148436A2-20230511-C00020
    Figure US20230148436A2-20230511-C00021
  • In a preferred embodiment, the present invention relates to small-molecule immune agonists of Formula (I), or isomers or pharmaceutically acceptable salts thereof, which are SZU-series compounds selected from the group consisting of (see Table 1 shown below for specific structural formulae): SZU-104, SZU-105, SZU-107, SZU-108, SZU-109, SZU-110, SZU-111, SZU-112, SZU-113, SZU-115, SZU-118, SZU-120, SZU-127, SZU-128, SZU-129, SZU-131, SZU-132, SZU-133, SZU-134, SZU-135, SZU-136, SZU-137, SZU-138, SZU-139, SZU-140, SZU-142, SZU-143, SZU-144, SZU-145, SZU-149, SZU-158, SZU-159, SZU-160, SZU-161, SZU-162, SZU-163, SZU-166 and SZU-171, or pharmaceutically acceptable salts thereof.
  • In the second aspect, the present invention relates to immune targeted compounds or pharmaceutically acceptable salts thereof, which are formed by covalently conjugating the small-molecule immune agonists or an addition product formed from the immune agonists and mercapto-containing polypeptides such as glutathione, with targeted drugs or antibodies, and represented by the following general formula:

  • (TLA)n-L-(Tar)m
  • wherein, TLA represents small-molecule immune agonists or their addition products with mercapto-containing polypeptides such as glutathione; Tar represents targeted drugs or antibodies; n and m each independently represent a numerical value ranging from 1 to 5; L represents a linker, which is linear or branched alkyl or polyethylene glycol chain, and both ends of L are covalently linked to TLA and/or Tar directly or via additional groups.
  • In a preferred embodiment, the small-molecule immune agonists comprise the small-molecule immune agonists as described in the first aspect of the present invention and compounds selected from the group consisting of:
  • Figure US20230148436A2-20230511-C00022
    Figure US20230148436A2-20230511-C00023
  • In a preferred embodiment, the small-molecule immune agonists are SZU-series compounds selected from the group consisting of (see Table 1 shown below for specific structural formulae): SZU-102, SZU-104, SZU-105, SZU-106, SZU-107, SZU-108, SZU-109, SZU-110, SZU-111, SZU-112, SZU-113, SZU-115, SZU-118, SZU-120, SZU-127, SZU-128, SZU-129, SZU-131, SZU-132, SZU-133, SZU-134, SZU-135, SZU-136, SZU-137, SZU-138, SZU-139, SZU-140, SZU-142, SZU-143, SZU-144, SZU-145, SZU-149, SZU-158, SZU-159, SZU-160, SZU-161, SZU-162, SZU-163, SZU-166, SZU-171, SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130, or pharmaceutically acceptable salts thereof.
  • In a preferred embodiment, the TLA represents compounds selected from the group consisting of (see Table 3 shown below for specific structural formulae): SZU-103-GSH, SZU-114-GSH, SZU-117-GSH, SZU-122-GSH, SZU-127-GSH, SZU-133-GSH, SZU-138-GSH and SZU-144-GSH.
  • In a preferred embodiment, the Tar represents at least one compound selected from the group consisting of: Dasatinib, Imatinib, Saracatinib, Ponatinib, Nilotinib, Danusertib, AT9283Degrasyn, Bafetinib, kw-2449, nyp-bhg712, dcc-2036, GZD824, GNF-2, PD173955, GNF-5, Bosutinib, Gefitinib, Erlotinib, Sunitinib, Ruxolitinib, Tofacitinib, Lapatinib, Vandetanib, Sorafenib, Sunitinib, Axitinib, Nintedanib, Regorafenib, Pazopanib, Lenvatinib, Crizotinib, Ceritinib, Cabozantinib, DWF, Afatinib, Ibrutinib, Niraparib, Palbociclib, B43, KU004, Foreinib, KRCA-0008, PF-06439015, PF-06463922, Canerlinib, GSA-1, GW2974, GW583340, W24002, CP-380736, D2667, Mubritinib, PD153035, PD168393, Pelitinib, PF-06459988, PF06672131, PF-6422899, PKI166, ReveromycinA, Tyrphostin1, tyrphostin23, lyrphostin51, Tyrphostin AG528, Tyrphostin AG658, Tyrphostin AG825, Tyrphostin AG835, Tyrphostin AG1478, Tyrphostin RG13022, Tyrphostin RG14620, B178, GSK1838705A, PD-161570, PD173074, SU-5402, Roslin2, Picropodophyllotoxin, PQ401, i-ome-tyrphostin, AG538, GNF5837, GW441756, Tyrphostin, AG879, DMPQ, jnj-10198409, PLX647, Trapidil, TyrphostinA9, Tyrhostin, AG370, Lestaurtinib, DMH4, Eldanamvcin, Genistein, Gw2580, HerbimycinA, Lavendustin, CMidostaurin, nvp-bhg712, PD158780, pd-166866, pf-0627334, PP2, RPL, SUl1274, SU5614, Symadex, Tyrphostin AG34, Tyrphostin AG974, Tyrphostin AG1007, UNC2881, Honokiol, SU1498, SKLB1002, cp-547632, jk-p3, KRN633, sc-1, ST638, SU 5416, Sulochrin, Tyrphostin SU1498, rociletinib, Dacomitinib, Tivantinib, Neratinib, Masitinib, Vatalanib, Icotinib, xl-184, osi-930, AB1010, Quizartinib, AZD9291, Tandutinib, HM61713, Brigantinib, Vemurafenib (plx-4032), Semaxanib, AZD2171, Crenolanib, Damnacanthal, Fostamatinib, Motesanib, Radotinib, osi-027, Linsitinib, BIX02189, PF-431396, PND-1186, PF-03814735, PF-431396, sirollmus, temsirolimus, everolimus, deforolimus, Zotarolimus, BEZ235, INK128, Omipalisib, AZD8055, MHY1485, PI-103, KU-0063794, ETP-46464, GDC0349, XL388, WYE-354, WYE-132, GiSK1059615, WAY-600, PF-04691502, wYE-687, PP121, BGT226, AZD2o014, PP242, CH5132799, P529, GDC0980, GDC-0994, XMD82, Ulixertinib, FR180204, SCH772984, Trametinib, PD184352, PD98059, Selumetinib, PD325901, U0126, Pimasertinib, tak-733, AZD8330, Binimetinib, PD318088, SL-327, Refametinib, gdc-0623, Cobimetinib, bl-847325, Adaphostin, GNF2, PPYA, aim-100, ASP 3026, LFM, Toceranib, JQ1, Niraparib, Fuzuopali, palbociclib, ARS-853, ARS-1620, Chlorazol-violet N (Direct violet N), miRNA-21, PreS, Zidovudine, Lenvatinib, LY-364947, ARS-1620 or Reparixin, EGFR inhibitors, TKI, BRD4 inhibitors, KRAS pathway-related target inhibitors, BRAF inhibitors, BTK Inhibitors, PARP inhibitors, PD-L1 inhibitors (including antibodies and small-molecules), PD-1 inhibitors (including antibodies and small-molecules), OX40 agonists (including antibodies and small-molecules), CD122 antibodies, CD3 antibodies, CD19 inhibitors, CD20 inhibitors, MUC1 inhibitors, MUC16 inhibitors, CDK4/6 inhibitors, TGF-0 inhibitors, CXCR inhibitors, CCL and CXCL chemokine inhibitors and miRNAs.
  • In a further preferred embodiment, the Tar represents at least one of the compounds selected from the group consisting of: TKI, EGFR inhibitors, KRAS inhibitors, BRAF inhibitors, BTK inhibitors, BRD4 inhibitors, PD-L1 inhibitors, PD-1 inhibitors, OX40 agonists, PARP inhibitors and CDK4/6 inhibitors.
  • In a preferred embodiment, L is
  • Figure US20230148436A2-20230511-C00024
  • In a preferred embodiment, the additional groups include amides, alkoxy bonds, alkylthio bonds, substituted amine groups, quaternary amine salts, heterocycles formed by Click reaction, amino acid groups that are dissociable by proteases, ureido or thiourea groups.
  • In a preferred embodiment, the immune targeted compounds or pharmaceutically acceptable salts thereof are SZU-series compounds selected from the group consisting of (see Table 4 shown below for specific structural formulae): SZU-116, SZU-119, SZU-124, SZU-125, SZU-146, SZU-147, SZU-168, SZU-169, SZU-174, SZU-175, SZU-176, SZU-177, SZU-178, SZU-179, SZU-180, SZU-181, SZU-158-PD-L1, SZU-158-OX40, SZU-158-PD-1 or SZU-136-miRNA21, or pharmaceutically acceptable salts thereof.
  • In the third aspect, the present invention relates to use of the small-molecule immune agonists of formula (I) or isomers or pharmaceutically acceptable salts thereof according to the first aspect of the present invention, compounds SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130 (see Table 2 shown below for specific structural formulae) and the immune targeted compounds or pharmaceutically acceptable salts thereof according to the second aspect of the present invention, in the manufacture of a medicament that participates in immunomodulation.
  • In the fourth aspect, the present invention relates to use of the small-molecule immune agonists of formula (I) or isomers or pharmaceutically acceptable salts thereof according to the first aspect of the present invention, compounds SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130 and the immune targeted compounds or pharmaceutically acceptable salts thereof according to the second aspect of the present invention, in the manufacture of a medicament as an anti-tumor or anti-viral drug or for eliminating target proteins.
  • The present invention further relates to use of a compound SZU-136-miRNA21 and analogues thereof in the manufacture of a medicament for immunologically targeting RNA, wherein the compound SZU-136-miRNA21 and analogues thereof are useful as small molecular RNAs for immune targeting; and the medicament may be present in various dosage forms, including solid preparations, liquid preparations, and conjugates formed from the medicament with various vehicles or crystalline hydrates of the medicament.
  • The present invention further relates to use of compounds SZU-179, SZU-180 and SZU-181 in the manufacture of a medicament as anti-AIDS and anti-hepatitis B agents, wherein the compounds SZU-179, SZU-180 and SZU-181 are useful as antiviral immune targeted compound.
  • The present invention further relates to use of compounds SZU-161 and SZU-162 for determining the conjugation degree of the small-molecule immune agonists conjugated with protein drugs and cells, wherein the compounds SZU-161 and SZU-162 are useful as small-molecule immune agonists with specific ultraviolet absorption.
  • The present invention further relates to use of compounds SZU-116 and SZU-124 in observing and studying the cellular absorption and distribution in vivo of proteins or targeted drugs, wherein compounds SZU-116 and SZU-124 are conjugated with proteins or targeted drugs, and are useful as compounds with optical tracking properties of chromogenic indicators for small-molecule immune agonists. After having been conjugated with proteins or targeted drugs, they can show the cellular absorption and distribution in vivo of proteins or targeted drugs in real time.
  • The present invention further relates to use of compounds SZU-114, SZU-117, SZU-122, SZU-127, SZU-133, SZU-138, and SZU-143 in the manufacture of a medicament with dual immune targeted functions, wherein the compounds SZU-114, SZU-117, SZU-122, SZU-127, SZU-133, SZU-138, and SZU-143 are conjugated with targeted drugs or antibodies.
  • Persons skilled in the art have surprisingly found that the small-molecule immune agonists of the present invention per se have immunostimulatory effects. Furthermore, it is well-known to the skilled persons that the combined application of TKI and TLR7 immune agonist will cause TKIs to antagonize the activation of TLR7 agonists on immunocytes, thereby inhibiting the production of immunostimulatory cytokines. However, by covalent conjugation of the TKIs as the targeted drugs with TLR7 small-molecule immune agonists, the inventors have surprisingly found that such covalently conjugated compounds not only maintain or improve the immunostimulatory effects of the small-molecule immune agonists, but also enhance the anti-tumor effects of the targeted drugs. The immune targeted compounds obtained by the method of the present invention are of great significance for the long-term and high-effective treatment of tumors.
  • The novel small-molecule immune agonists that have immunostimulatory effects and can be conjugated with targeted drugs to form immune targeted compounds with targeting action as described above are shown in Table 1 below:
  • TABLE 1
    Serial Molecular
    No. Structural Formula weight
    SZU- 104
    Figure US20230148436A2-20230511-C00025
         		632.74
    SZU- 105
    Figure US20230148436A2-20230511-C00026
         		532.62
    SZU- 107
    Figure US20230148436A2-20230511-C00027
         		647.58
    SZU- 112
    Figure US20230148436A2-20230511-C00028
         		794.87
    SZU- 113
    Figure US20230148436A2-20230511-C00029
         		572.62
    SZU- 108
    Figure US20230148436A2-20230511-C00030
         		563.63
    SZU- 109
    Figure US20230148436A2-20230511-C00031
         		607.68
    SZU- 110
    Figure US20230148436A2-20230511-C00032
         		518.53
    SZU- 111
    Figure US20230148436A2-20230511-C00033
         		562.58
    SZU- 144
    Figure US20230148436A2-20230511-C00034
         		442.43
    SZU- 133
    Figure US20230148436A2-20230511-C00035
         		442.42
    SZU- 138
    Figure US20230148436A2-20230511-C00036
         		456.46
    SZU- 134
    Figure US20230148436A2-20230511-C00037
         		458.48
    SZU- 135
    Figure US20230148436A2-20230511-C00038
         		430.42
    SZU- 158
    Figure US20230148436A2-20230511-C00039
         		561.63
    SZU- 159
    Figure US20230148436A2-20230511-C00040
         		547.63
    SZU- 161
    Figure US20230148436A2-20230511-C00041
         		707.80
    SZU- 162
    Figure US20230148436A2-20230511-C00042
         		694.77
    SZU- 115
    Figure US20230148436A2-20230511-C00043
         		581.63
    SZU- 160
    Figure US20230148436A2-20230511-C00044
         		598.62
    SZU- 127
    Figure US20230148436A2-20230511-C00045
         		626.80
    SZU- 145
    Figure US20230148436A2-20230511-C00046
         		617.62
    SZU- 143
    Figure US20230148436A2-20230511-C00047
         		456.43
    SZU- 149
    Figure US20230148436A2-20230511-C00048
         		358.35
    SZU- 163
    Figure US20230148436A2-20230511-C00049
         		328.37
    SZU- 166
    Figure US20230148436A2-20230511-C00050
         		354.36
    SZU- 139
    Figure US20230148436A2-20230511-C00051
         		653.70
    SZU- 128
    Figure US20230148436A2-20230511-C00052
         		526.60
    SZU- 129
    Figure US20230148436A2-20230511-C00053
         		529.62
    SZU- 131
    Figure US20230148436A2-20230511-C00054
         		554.61
    SZU- 132
    Figure US20230148436A2-20230511-C00055
         		497.56
    SZU- 120
    Figure US20230148436A2-20230511-C00056
         		513.56
    SZU- 118
    Figure US20230148436A2-20230511-C00057
         		459.46
    SZU- 136
    Figure US20230148436A2-20230511-C00058
         		424.45
    SZU- 142
    Figure US20230148436A2-20230511-C00059
         		344.37
    SZU-
    140
    SZU- 137
    Figure US20230148436A2-20230511-C00060
    SZU- 171
    Figure US20230148436A2-20230511-C00061
  • Other small-molecule immune agonists that have immunostimulatory effects and can be conjugated with targeted drugs to form immune targeted compounds with targeting action are shown in Table 2 below:
  • TABLE 2
    Serial Molecular
    No. Structural Formula weight
    SZU- 101
    Figure US20230148436A2-20230511-C00062
    SZU-
    102
    SZU- 103
    Figure US20230148436A2-20230511-C00063
    SZU- 114
    Figure US20230148436A2-20230511-C00064
         		455.52
    SZU- 117
    Figure US20230148436A2-20230511-C00065
         		566.62
    SZU- 122
    Figure US20230148436A2-20230511-C00066
         		398.42
    SZU- 130
    Figure US20230148436A2-20230511-C00067
         		512.57
    SZU- 106
    Figure US20230148436A2-20230511-C00068
         		603.63
  • Among such compounds, the structural formulae of compounds SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130 and their preparation processes have been disclosed in the Chinese Patent Application CN107281483A; and the structural formulae of compounds SZU-102 and SZU-106 and their preparation processes have been disclosed in the Chinese Patent Application CN106267188A, wherein compound 1 corresponds to compound SZU-102 and compound 19 corresponds to compound SZU-106.
  • Some small-molecule immune agonists listed in Tables 1 and 2 can form addition products with mercapto-containing polypeptides such as glutathione (GSH), which can target glutathione transferase, result in improved therapeutic efficacy on diseases by acting synergistically with chemotherapy drugs, irreversible TKI targeted drugs and other irreversible targeted drugs, and eliminate drug resistance, while activating immunological effects. One of the addition products, SZU-117-GSH formed by reacting SZU-117 with glutathione via a synthesis process as shown in the following scheme, is illustrated as an example. Examples of the addition products formed from the small-molecule immune agonists and GSH are shown in Table 3:
  • Figure US20230148436A2-20230511-C00069
  • TABLE 3
    SZU- 103- GSH
    Figure US20230148436A2-20230511-C00070
    SZU- 114- GSH
    Figure US20230148436A2-20230511-C00071
    SZU- 117- GSH
    Figure US20230148436A2-20230511-C00072
    SZU- 122- GSH
    Figure US20230148436A2-20230511-C00073
    SZU- 127- GSH
    Figure US20230148436A2-20230511-C00074
    SZU- 133- GSH
    Figure US20230148436A2-20230511-C00075
    SZU- 138- GSH
    Figure US20230148436A2-20230511-C00076
    SZU- 144- GSH
    Figure US20230148436A2-20230511-C00077
  • Other mercapto-containing compounds include, for example, 2-methylamino ethanethiol and
  • Figure US20230148436A2-20230511-C00078
  • The small-molecule immune agonists listed in Tables 1 and 2 and the compounds formed from some small-molecule immune agonists and GSH listed in Table 3 can be conjugated with targeted drugs to give bi-functional immune targeted compounds with immune targeting effects. The immune targeted compounds obtained are shown in Table 4 below:
  • TABLE 4
    Serial Molecular
    No. Structural Formula weight
    SZU- 116
    Figure US20230148436A2-20230511-C00079
         		1393.63
    SZU- 119
    Figure US20230148436A2-20230511-C00080
         		1001.56
    SZU- 124
    Figure US20230148436A2-20230511-C00081
         		1189.27
    SZU- 125
    Figure US20230148436A2-20230511-C00082
         		 801.95
    SZU- 146
    Figure US20230148436A2-20230511-C00083
         		 618.58
    SZU- 147
    Figure US20230148436A2-20230511-C00084
         		 746.83
    SZU- 169
    Figure US20230148436A2-20230511-C00085
         		1565.61
    SZU- 174
    Figure US20230148436A2-20230511-C00086
         		1148.07
    SZU- 175
    Figure US20230148436A2-20230511-C00087
         		1094.67
    SZU- 168
    Figure US20230148436A2-20230511-C00088
         		 905.39
    SZU- 176
    Figure US20230148436A2-20230511-C00089
         		 869.97
    SZU- 177
    Figure US20230148436A2-20230511-C00090
         		 992.05
    SZU- 179
    Figure US20230148436A2-20230511-C00091
         		 848.86
    SZU- 158- PD- L1
    Figure US20230148436A2-20230511-C00092
    SZU- 158- OX40
    Figure US20230148436A2-20230511-C00093
    SZU- 158- PD-1
    Figure US20230148436A2-20230511-C00094
    SZU- 178
    Figure US20230148436A2-20230511-C00095
    SZU- 180
    Figure US20230148436A2-20230511-C00096
    SZU- 181
    Figure US20230148436A2-20230511-C00097
         		3512.80
    SZU- 136- miR NA21
    Figure US20230148436A2-20230511-C00098
  • Small-molecule immune agonists are conjugated with targeted drugs to form immune targeted compounds via synthesis schemes shown below:
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme I:
  • Figure US20230148436A2-20230511-C00099
  • Small-molecule immune agonists (using SZU-137 as an example) may be conjugated with targeted drugs via Synthesis Scheme II:
  • Figure US20230148436A2-20230511-C00100
    Figure US20230148436A2-20230511-C00101
  • The same process as described above may also be applied to conjugating of SZU-144 or SZU-133 with targeted drugs to form immune targeted compounds. The linker may also be a simple linear alkane or other linkers replaced as desired.
  • Carboxyl-containing small-molecule immune agonists (such as SZU-138) can be conjugated with antibodies through addition reactions and conjugation reactions to form soluble, functional protein polypeptide conjugates, which can be formed into salts owing to the presence of the dimethylamino group, so that they can provide ionized positive charges, and thus are advantageous for increasing the therapeutic efficacy and water solubility, thereby solving the solubility problem of the conjugated products.
  • Furthermore, the carboxyl-containing small-molecule immune agonists as described above can also be conjugated with two different targeted drugs to form dual-targeted immune compounds.
  • Small-molecule immune agonists (using SZU-171 as an example) may be conjugated with targeted drugs via Synthesis Scheme III:
  • Figure US20230148436A2-20230511-C00102
  • Products formed from small-molecule immune agonists and glutathione (using SZU-114-GSH as an example) can also be conjugated with two different targeted drugs to enhance the therapeutic efficacy.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme IV:
  • Figure US20230148436A2-20230511-C00103
  • As used herein, the targeted drugs can either be various TKI agents and tumor targeted drugs, or antibodies such as CD3 antibody, PD-1 antibody, OX40 antibody, CD19 antibody, CD20 antibody, HER2 antibody, MUC1 antibody and antibodies against various tumor proteins and pathogens, wherein the linker may be alkane or alkoxy chains, and can be adjusted depending on the requirements for optimizing dissolution and metabolism.
  • Dual conjugated immune-targeted compounds as described above have the following advantages:
      • Being capable of forming two positive amino ionic groups (amino salts —HN+— and —NH3 +);
      • High solubility;
      • Dual targets, which can be adjusted and optimized if necessary, for example, with target 1 being CD3 antibody or CD122 antibody targeting T cells, and target 2 being HER2 inhibitors targeting cancer cells;
      • promoting infiltration of T cells into the tumor microenvironment;
      • enhancing the anti-tumor effects while activating immune TLR7 receptors; and
      • forming immune targeted memory.
  • Small-molecule immune agonists (using SZU-166 as an example, which is a unique isocyanate small-molecule immune agonist with the merit of mild quantitative conjugate addition with amino compounds) may be conjugated with targeted drugs via Synthesis Scheme V:
  • Figure US20230148436A2-20230511-C00104
  • Compounds SZU-115 and SZU-160 are novel small-molecule immune agonists with Click reactive groups that mainly have two functions including: 1) after having been conjugated with macromolecules and biologically active substances, being useful in optical tracking of the conjugated product by linking to a chromogenic indicator via Click reaction; for example, when they have conjugated with proteins or targeted drugs, the cellular absorption and distribution in vivo of the proteins and targeted drugs are observed.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme VI:
  • Figure US20230148436A2-20230511-C00105
  • 2) SZU-115 can also be simultaneously linked to antibodies and targeted drugs, thereby leading to reduced off-target effects of the antibodies, enhanced efficacy of the targeted drugs and activated antitumor immunity:
  • Figure US20230148436A2-20230511-C00106
  • Compounds SZU-136 and SZU-160 have similar action mechanisms and application effects.
  • Compound SZU-139 in the present invention is prepared by Click reaction of SZU-136 with ribose:
    • Conjugation of Immune Agonist with Ribose
  • Figure US20230148436A2-20230511-C00107
  • The same process can also be applied for conjugating small-molecule immune agonists with small-molecule RNAs (miRNAs). For example, miRNA21 (microRNA21) plays an important role in generation and development of tumor (Cell Death Dis. 2018 Feb. 13; 9(2):219. doi: 10.1038/s41419-017-0243-9; Proc Natl Acad Sci S U.S.A. 2015 Jun. 30; 112(26): E3355-64. doi: 10.1073/pnas.1504630112; Cancer Lett. 2017 May 1; 393: 86-93.doi:10.1016/j.canlet.2017.02.019; https://en.wikipedia.org/wiki/MIRN21). The application of small-molecule immune agonists conjugated with pathogenic miRNAs can guide the immune system to block the occurrence of diseases at source, the changes of related proteins and the onset of related diseases.
  • Small-molecule immune agonists may be conjugated with targeted drugs via Synthesis Scheme VII:
  • Figure US20230148436A2-20230511-C00108
  • Synthesis of azide miRNA was described in the literature (Bioconjug Chem. 2003 May-Jun.; 14(3):697-701):
  • Figure US20230148436A2-20230511-C00109
  • The alkyne immune agonist can be replaced with SZU-115. The carboxyl group of SZU-115 can be conjugated with Argonaute to form a conjugate, which interrupts the formation of a conjugate of miRNA and Argonaute, and is directed into the endosome of immunocytes under the action of TLR7 agonist, thus resulting in immunological elimination and memory:
  • Figure US20230148436A2-20230511-C00110
  • Recent studies have confirmed that pathogenic miRNAs released by tumor exosomes present an important cause of drug resistance and tumor metastasis (Proc, Natl, Acad, Sci, U, A, 2018, February, 12.pii:201717363.doi:10.1073/pnas.1717363115, J Clin Invest. 2016 Apr. 1; 126(4): 1163-72.doi: 10.1172/JCI81130); self-miRNAs generated in human bodies, especially those released from cancer cells, are generally incapable of inducing immunity. Although the target miRNAs can be specifically recognized by exogenous antisense oligonucleotide RNAs, most of them will be cleared or degraded rapidly in vivo owing to their immunogenicity. By using the above process, it is possible to transform pathogenic self-miRNAs into immunogenic miRNAs that can be recognized by innate immunity. The same process can also be applied to other pathogenic miRNAs such as miRNA30c.
  • An important role of miRNA is to guide the corresponding Argonaute protein to initiate target mRNA cleavage or transcriptional suppression (https://en.wikipedia.org/wiki/Argonaute). Therefore, one of the innovative effects achieved by the TLR7 immune agonist conjugated with self-miRNAs in human bodies lies in immune targeted interference in the binding to the Argonaute protein, thereby eliminating the related pathogenicity. This effect is schematically shown in FIG. 69 . In summary, the inventors have found a series of small-molecule immune agonists, which can be conjugated with targeted drugs to form novel immune targeted compounds with immunostimulatory function, thereby resulting in increased in vitro and in vivo immunostimulatory effects of the targeted drugs and improved efficacy against tumors and other diseases, conferring dual functions to the targeted drugs (bi-functional targeted drugs). Such an enhanced effect is resulted from the synergistic action of two functions: inclusion of antitumor immune factors (such as IFN-γ) and inhibition on the pathogenic targeted sites. Thus, the present invention is directed to novel high-effective targeted drugs with a combination of the two functions.
  • The immune targeted compounds according to the present invention act appropriately by eliminating pathogenic targets (proteins) via immune pathway as shown in FIG. 70 . Immune targeted compounds formed by conjugating small-molecule immune agonists with targeted drugs are guided by the targeted drugs to the pathogenic proteins, thereby resulting in the death of the targeted cells and release of the pathogenic proteins. Further, they are guided by the immune agonists to antigen presenting cells (such as DC), in which the pathogenic proteins are cleaved, leading to degradation of these proteins. The immune targeted compounds formed by conjugating small-molecule immune agonists with targeted drugs (such as TKIs) in the present invention can overcome drug resistance and improve therapeutic efficacy.
  • Lysine-derived compounds SZU-104, SZU-105, SZU-112 and SZU-113 that are useful as acidic small-molecule immune agonists are amino acid derivatives having the activity of immune agonists, and can achieve the purpose of site-specific conjugation by inserting polypeptides (or antibodies) at specific sites as desired in the preparation of immunogenic polypeptides:
  • Figure US20230148436A2-20230511-C00111
  • SZU-161 and SZU-162, as small-molecule immune agonists suitable for conjugating with protein drugs, have a specific ultraviolet absorption peak (342 nM), and can be used typically for measuring the conjugation degree of small-molecule immune agonists conjugated with proteins drugs:
  • Figure US20230148436A2-20230511-C00112
  • The measurement method is described as follows: 1) SZU-161 was measured for the intensity of optical absorption at 342 nM under different concentrations; standard curves of concentration and absorption were plotted to obtain the concentration-absorption relationship formula; and 2) after conjugation with protein, the conjugated protein was measured for the absorption at 342 nM under certain concentration; the resulting value was placed into the relationship formula to readily obtain the relative concentration of SZU-161 in the conjugate (i.e., conjugation degree). SZU-162 may be applied to achieve the same effect.
    • BRIEF DESCRIPTION OF THE DRAWINGS
  • The “immune cytokines” mentioned in the following figures refer to cytokines (including IFN-γ, IL-12, IL-6, and TNF-α), and their concentration levels can be used to indicate the levels of immunocyte responsiveness stimulated by the compound.
  • FIG. 1 shows the stimulatory effects of SZU-GSH-series compounds on immune cytokine (IL-6).
  • FIG. 2 shows the distribution of fluorescence-labeled SZU-116 in cells.
  • FIG. 3 shows the stimulatory effects of SZU-102, 103, 106, 107, 108, 110, 111, 114, 115, 116, 117, and 118 on immune cytokine (IFN-γ), wherein SZU-101 and R848, as the standard control, are internationally-accepted, standard small-molecule immune agonists with immunostimulatory function (the same below).
  • FIG. 4 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (IFN-γ), wherein “con” is PBS blank control (the same below).
  • FIG. 5 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (IL-12).
  • FIG. 6 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (TNF-α).
  • FIG. 7 shows the stimulatory effects of SZU-143, 144, 146, 147, and 149 on immune cytokine (IL-6).
  • FIG. 8 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokine (IFN-γ).
  • FIG. 9 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokine (IL-12).
  • FIG. 10 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokine (TNF-α).
  • FIG. 11 shows the stimulatory effects of SZU-145, 158, 104, 109, 112, and 124 on immune cytokines (IL-6).
  • FIG. 12 shows the stimulatory effects of SZU-125 on several immune cytokines.
  • FIG. 13 shows the stimulatory effects of SZU-105 on immune cytokine (IFN-γ).
  • FIG. 14 shows the stimulatory effects of SZU-105 on immune cytokine (IL-12).
  • FIG. 15 shows the stimulatory effects of SZU-113 on immune cytokine (IFN-γ).
  • FIG. 16 shows the stimulatory effects of SZU-113 on immune cytokine (IL-12).
  • FIG. 17 shows the stimulatory effects of SZU-119 on immune cytokine (IL-12).
  • FIG. 18 shows the stimulatory effects of SZU-119 on immune cytokine (IL-6).
  • FIG. 19 shows the stimulatory effects of SZU-120 on immune cytokine (IFN-γ).
  • FIG. 20 shows the stimulatory effects of SZU-120 on immune cytokine (IL-6).
  • FIGS. 21-54 show the stimulatory effects of the small-molecule immune agonists SZU-122 and 127-142 according to the present invention on immune cytokines, respectively.
  • FIG. 55 shows the stimulatory effects of SZU-102, 119, 125, 146, 147, 169, 174, 175, 168, 176, 177, and 179 on immunocytes to produce IFN-γ.
  • FIG. 56 shows the stimulatory effects of SZU-102, 119, 125, 146, 147, 169, 174, 175, 168, 176, 177, and 179 on immunocytes to produce IL-12.
  • FIG. 57 shows the anti-tumor (CT26) activity of novel immune targeted compounds in vivo: Veh (untreated control), A (SZU-177), B (SZU-175), C (SZU-174), D (SZU-147), and E (SZU-176).
  • FIG. 58 shows the anti-tumor (B16) activity of novel immune targeted compounds in vivo: Veh (untreated control), G (PD-L1 antibody), H (SZU-178), I (SZU-158-PD-L1), J (SZU-158-OX40), K (PD-1 antibody), and L (SZU-158-PD-1).
  • FIG. 59 shows the inhibition of JQ1 on the activity of SZU-101 as a TLR7 agonist; wherein SZU-119 retains the activity of TLR7 agonist for stimulating immune cytokine.
  • FIG. 60 shows the inhibition of JQ1 on the activity of SZU-101 as a TLR7 agonist, wherein SZU-119 retains the activity of TLR7 agonist for stimulating immune cytokine.
  • FIG. 61 shows that SZU-119 retains the inhibitory activity of JQ1 against CD274 (PD-L1) on B16 cells, wherein “control” represents blank control.
  • FIG. 62 shows the stimulatory effects of SZU-163 and SZU-166 on immunocytes to produce IFN-γ.
  • FIG. 63 shows the stimulatory effects of SZU-180 and SZU-136-miRNA21 on immunocytes to produce IFN-γ.
  • FIG. 64 shows the anti-tumor (4T1) survival rates of PD-1 antibody, PD-L1 antibody, OX40 antibody and their SZU-158 conjugates and SZU-178.
  • FIG. 65 shows the stimulatory effects of PD-1 antibody, PD-L1 antibody, OX40 antibody, and their SZU-158 conjugates and SZU-178 on immunocytes to produce IFN-γ.
  • FIG. 66 shows the stimulatory effects of SZU-181 on immunocytes to produce IFN-γ.
  • FIG. 67 shows the stimulatory effects of SZU-106, SZU-158, and SZU-160 on immunocytes to produce IFN-γ, wherein “control” represents PBS blank control.
  • FIG. 68 shows the stimulatory effects of SZU-106, SZU-158, and SZU-160 on immunocytes to produce IL-6, where “control” represents blank control.
  • FIG. 69 is a schematic diagram showing the functioning mechanism of TLR7 small-molecule immune agonist conjugated with self-miRNAs in human bodies for immune-targeted interference in the binding of agonist to protein, thereby eliminating protein-related pathogenicity.
  • FIG. 70 is a schematic diagram showing the functioning mechanism of the immune targeted compound of the present invention for eliminating pathogenic targets (proteins) via immune pathway.
    •  DETAILED DESCRIPTION OF THE INVENTION
  • Processes for the synthesis of the related compounds of the present invention are exemplified as follows. The exemplified synthesis processes could inspire those skilled in the art to achieve the synthesis of the novel compounds as described herein. However, synthesis of the novel compounds of the present invention is not limited to these exemplified synthesis processes.
    • PREPARATION EXAMPLES
  • Figure US20230148436A2-20230511-C00113
  • 1 g of SZU-101, 311 mg of N-hydroxysuccinimide (NHS) and 518 mg of dichloroethane (EDC) were dissolved in 12 mL of anhydrous N,N-dimethylformamide (DMF). The mixture was reacted at room temperature for 2 hours under the protection of nitrogen atmosphere. The reaction was monitored by Thin Layer Chromatography (TLC). When the reaction was completed, the reaction solution was poured into water. The precipitate was filtered off and dried to give a crude SZU-009T, which was directly reacted in the next step.
  • 500 mg of SZU-009T and 339 mg of Fmoc-D-lysine were dissolved in 10 mL of anhydrous DMF. The mixture was stirred at room temperature for 1 hour. After the reaction was completed, the mixture was admixed with water, and precipitated solids were filtered off with suction. The residue was purified by column chromatography (chloroform:methanol=3:1) to give a white solid (576 mg) with a yield of 78.9%, ESI-MS: m/z=795.3 [M+H]+.
  • Figure US20230148436A2-20230511-C00114
  • 1 g of SZU-101 was dissolved in 20-fold weight of anhydrous DMSO. Equimoles of compounds 1), 2) and 3) were added successively. The reactants were mixed under stirring at room temperature for 12 hours, and added with 10-fold volume of water based on the reaction volume. The mixture was centrifuged to give a precipitate as a crude product, which was added into 20 mL of saturated aqueous sodium carbonate solution. The resulting mixture was stirred for 10 minutes and filtered. The filtrate was adjusted to a pH of 3 with concentrated hydrochloric acid. A pure product SZU-107 was precipitated out and dried under vacuum, with a yield of 65%. ESI-MS: m/z=647.6865 [M+H]+.
  • Figure US20230148436A2-20230511-C00115
  • 50 mg of SZU-112 was dissolved in 1 mL of diethylamine. The mixture was stirred at room temperature for 1 hour. The progress of the reaction was monitored by TLC. After the reaction was completed, diethylamine was removed under reduced pressure. The residue was washed with the addition of 1 mL of dichloromethane, then centrifuged at 10,000 rpm for 5 minutes. The supernatant was removed. The precipitate was washed again and dried under reduced pressure to give 32 mg of a white solid with a yield of 90%, ESI-MS: m/z=573.2 [M+H]+.
  • Figure US20230148436A2-20230511-C00116
  • 108 mg of SZU-009T was dissolved in 2 mL of anhydrous DMF. 31 mg of SZU-013T and 100 μL of N,N-diisopropylethylamine (DIEA) were added successively. The mixture was stirred at room temperature overnight. The progress of the reaction was monitored by LC-MS. After the reaction was completed, the residue was purified by preparative liquid chromatography to give 65 mg of a white solid with a yield of 56%. ESI-MS: m/z=582.2 [M+H]+.
  • Figure US20230148436A2-20230511-C00117
  • 264 mg of SZU-017T, 324 mg of HBTU and 119 mg of 1-hydroxybenzotriazole (HOBt) was dissolved in 5 mL of anhydrous DMF. 256 μL of DIEA was added dropwise. The mixture was stirred at room temperature for 10 minutes. 5 mL of SZU-020T (230 mg) in anhydrous DMF was added and the resulting mixture was stirred at room temperature overnight. The reaction solution was poured into 100 mL of 5% HCl, and extracted with 30 mL of dichloromethane three times. The organic layers were combined and washed with saturated sodium bicarbonate solution, 5% HCl, and saturated sodium chloride solution, respectively, and dried over anhydrous sodium sulfate. The solvent was removed under reduced pressure, and the residue was purified by column chromatography (dichloromethane:methanol=50:1) to give 180 mg of a purple-red semi-solid SZU-021T with a yield of 51.6%.
  • 28 mg of SZU-021T was dissolved in 0.1 mL of dimethyl sulfoxide (DMSO) and admixed with 0.5 mL of aqueous solution of tris(2-carboxyethyl)phosphine (TCEP) (97 mg), 0.1 mL of TBTA (18 mg) in DMSO and 0.5 mL of aqueous solution of anhydrous copper sulfate (54 mg). 20 mg of SZU-115 in 2 mL of methanol/water mixed solution was added. The mixture was stirred at room temperature for 2 hours and purified by column chromatography to give 31 mg of a purple-red solid with a yield of 66%. ESI-MS: m/z=1393.7 [M+H]+.
  • Figure US20230148436A2-20230511-C00118
    Figure US20230148436A2-20230511-C00119
  • 667 mg of SZU-009T was dissolved in 2 mL of anhydrous DMF. 300 mg of SZU-020T was added and the mixture was stirred at room temperature overnight. After the reaction was completed, the reaction solution was poured into water. A white solid was precipitated out and dried to give a crude SZU-030T.
  • The SZU-030T obtained in the above step was dissolved in 10 mL of anhydrous tetrahydrofuran (THF). 484 mg of triphenylphosphine was added under an ice bath. The mixture was transferred to room temperature and continued to react under stirring overnight. The progress of the reaction was monitored by LC-MS. After the reaction was completed, 20 mL of water was added. The mixture was stirred for another 30 minutes, then filtered off with suction under reduced pressure. The residue was washed with water twice and dried to give 625 mg of SZU-031 T with a yield of 79% over the two steps.
  • 71 mg of SZU-031T, 42 mg of (+)-JQ-1-COOH (JQ1, active isomer), 46 mg of HBTU were dissolved in 2 mL anhydrous DMF. Catalytic amount of 4-dimethylaminopyridine (DMAP) and N,N-diisopropylethylamine (DIPEA) were added. The mixture was stirred at room temperature for 24 hours. The progress of the reaction was monitored by LC-MS. The residue was purified by preparative liquid chromatography and lyophilized to give 27 mg of a light yellow solid with a yield of 27%. ESI-MS: m/z=1001.2 [M+H]+.
  • Figure US20230148436A2-20230511-C00120
  • SZU-124 was prepared by the same process as SZU-116, except that SZU-017T was replaced with FITC, and a grass green semi-solid was obtained. The yield over the two steps was 31%. ESI-MS: m/z=1199.5 [M+H]+.
  • Figure US20230148436A2-20230511-C00121
  • 34 mg of SZU-008T was dissolved in 500 μL of anhydrous DMF. 53 mg of PD-1/PD-L1 inhibitor (CAS #:1675203-84-5), 43 mg of HBTU, 42 μL of triethylamine (TEA) and catalytic amount of DMAP were added successively. The mixture was stirred at room temperature for 5 hours. After the reaction was completed, the residue was purified by preparative liquid chromatography and lyophilized to give 45 mg of a white solid with a yield of 56%. ESI-MS: m/z=802.3 [M+H]+.
  • Figure US20230148436A2-20230511-C00122
  • SZU-127 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with retinoic acid, and a yellow solid was obtained. ESI-MS: m/z=627.4[M+H]+.
  • Figure US20230148436A2-20230511-C00123
  • SZU-134 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with glutaric anhydride; and a white solid was obtained. ESI-MS: m/z=459.1[M+H]+.
  • Figure US20230148436A2-20230511-C00124
  • SZU-138 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with itaconic anhydride; and a white solid was obtained. ESI-MS: m/z=457.1[M+H]+.
  • Figure US20230148436A2-20230511-C00125
  • SZU-140 was prepared by the same process as SZU-125, except that the PD-1/PD-L1 inhibitor was replaced with acetic anhydride; and a white solid was obtained. ESI-MS: m/z=387.1[M+H]+.
  • Figure US20230148436A2-20230511-C00126
  • SZU-103 and an equivalent amount of glutathione (Glutathione, GSH) were dissolved in anhydrous DMF. The mixture was stirred at room temperature overnight. The progress of the reaction was monitored by LC-MS. After the reaction was completed, the residue was purified by preparative liquid chromatography to give a white solid with a yield of 85%. ESI-MS: m/z=936.2 [M+H]+.
  • Figure US20230148436A2-20230511-C00127
  • 5 g of SZU-005T was placed in a round-bottom flask. Concentrated hydrochloric acid was added. The mixture was reacted for 12 hours. After the reaction was completed, the resulting mixture was adjusted to a pH of 3-4 with sodium hydroxide, and 4.56 g of a white solid was precipitated out, with the yield being 95%. ESI-MS: m/z=345.2[M+H]+.
  • Figure US20230148436A2-20230511-C00128
  • 20 mg of SZU-138 was dissolved in 2 mL of anhydrous DMF. 8.7 mg of D-glucosamine, 9.2 mg of EDC, and 6.5 mg of HOBt were added successively. The mixture was stirred at room temperature for 2 hours. After the reaction was completed, the residue was purified by preparative liquid chromatography to give 12 mg of a white solid, with the yield being 44%, ESI-MS: m/z=618.2 [M+H]+.
  • Figure US20230148436A2-20230511-C00129
  • SZU-114-GSH was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-114; and a white solid was obtained. ESI-MS: m/z=763.3[M+H]+.
  • Figure US20230148436A2-20230511-C00130
  • SZU-117-GSH was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-117; and a white solid was obtained. ESI-MS: m/z=874.3[M+H]+.
  • Figure US20230148436A2-20230511-C00131
  • SZU-158 was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-138; glutathione was replaced with 2-(dimethylamino)ethane-1-thiol; and white solid was obtained. ESI-MS: m/z=562.2[M+H]+. SZU-159 can be prepared by the same process, except that SZU-138 was replaced with SZU-144, which was reacted with 2-(dimethylamino)ethane-1-thiol at room temperature for 24 hours to give SZU-159.
  • Figure US20230148436A2-20230511-C00132
  • SZU-161 was prepared by the same process as SZU-103-GSH, except that SZU-103 was replaced with SZU-138; glutathione was replaced with SZU-033T; and a white solid was obtained. ESI-MS: m/z=708.4[M+H]+.
  • SZU-162
  • SZU-162 was prepared by the same process as SZU-161. ESI-MS: m/z=695.81[M+H]+.
  • Figure US20230148436A2-20230511-C00133
  • SZU-160 was prepared by the same process as SZU-115, except that SZU-013T was replaced with SZU-032T; and a white solid was obtained. ESI-MS: m/z=599.3[M+H]+.
  • Figure US20230148436A2-20230511-C00134
  • HBTU, DMAP and TEA were added to the anhydrous DMF solution of TL-008 at 0° C. The mixture was stirred at room temperature for 0.5 hour. Then trans-4-dimethylaminocrotonic acid hydrochloride was added at 0° C. The resulting mixture was reacted at room temperature for 24 hours. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=456.6 [M+H]+).
  • Figure US20230148436A2-20230511-C00135
  • Synthesis of SZU-001S:
  • 1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride (EDCI), HOBt and DIPEA were added to the anhydrous DMF solution of SZU-101 at 0° C. The mixture was stirred at room temperature for 0.5 hour. Then N-Boc piperazine was added at 0° C. The resulting mixture was reacted at room temperature for 24 hours. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=613.4 [M+H]+).
  • Synthesis of SZU-130:
  • Trifluoroacetic acid was added dropwise to the dichloromethane solution of SZU-001S. After stirring for 2 hours, the reaction solution was concentrated under reduced pressure. Then 5 mL ethanol and 2N HCl solution were added to give a hydrochloride salt. After the solvent was removed under reduced pressure, diethyl ether or dichloromethane was added, and then solvent was removed again under reduced pressure. This procedure was repeated several times to give a pure solid product (ESI-MS: m/z=513.5 [M+H]+).
  • Synthesis of SZU-117:
  • TEA and acryloyl chloride were added successively to the anhydrous DMF solution of SZU-130 at 0° C. After stirring at room temperature for 2 h, the reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=567.7[M+H]+).
  • Figure US20230148436A2-20230511-C00136
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 min, DIPEA was added at 0° C. The mixture was reacted at room temperature for 0.5 hour, and then hydroxylamine hydrochloride was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=460.4 [M+H]+).
  • Figure US20230148436A2-20230511-C00137
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 minutes, DIPEA was added at 0° C. The mixture was reacted at room temperature for 0.5 hour, and then morpholine was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=514.5[M+H]+).
  • Figure US20230148436A2-20230511-C00138
  • TEA and acryloyl chloride were added successively to the anhydrous DMF solution of SZU-142 at 0° C. After stirring at room temperature for 2 hours, the reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=399.4[M+H]+).
  • Figure US20230148436A2-20230511-C00139
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 minutes, DIPEA was added at 0° C. The mixture was reacted at room temperature for 0.5 hour, and then 1-methylpiperazine was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=527.7 [M+H]+).
  • Figure US20230148436A2-20230511-C00140
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 min, DIPEA was added at 0° C. The mixture was reacted at room temperature for 0.5 hour, and then thiomorpholine was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=530.6 [M+H]+).
  • Figure US20230148436A2-20230511-C00141
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 min, DIPEA was added at 0° C. The mixture was reacted at room temperature for 0.5 hour, and then 1-acetylpiperazine was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=555.6 [M+H]+).
  • Figure US20230148436A2-20230511-C00142
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 min, DIPEA was added at 0° C. The mixture was reacted at room temperature for 0.5 hour, and then pyrrolidine was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=498.5 [M+H]+).
  • Figure US20230148436A2-20230511-C00143
  • Maleic anhydride was added to the anhydrous DMF solution of SZU-142. After the resulting solution was reacted overnight, the reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=443.3 [M+H]+).
  • Figure US20230148436A2-20230511-C00144
  • EDCI and HOBt were added to the anhydrous DMF solution of monomethyl fumarate. After stirring for 15 min, DIPEA was added at 0° C. The resulting solution was reacted at room temperature for 0.5 hour, and then SZU-142 was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by high performance liquid chromatography (HPLC) to obtain a pure product (ESI-MS: m/z=457.4 [M+H]+).
  • Figure US20230148436A2-20230511-C00145
  • Sodium hydroxide (1.1 equivalent amount) was added to the methanol/water (1:1) mixed solution of SZU-143. After stirring at room temperature for 8 hours, the reaction solution was neutralized with 1N HCl solution after the reaction was completed, and then filtered off with suction and washed with water to give a crude product, which was purified by high performance liquid chromatography (HPLC) to obtain a pure product (ESI-MS: m/z=443.5[M+H]+).
  • Figure US20230148436A2-20230511-C00146
  • EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 minutes, DIPEA was added at 0° C. The resulting solution was reacted at room temperature for 0.5 hour, and then 3-(trifluoromethyl)-5,6,7,8-tetrahydro-[1,2,4]triazolo[4,3-a]pyrazine (intermediate of Fuzuopali) was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by liquid phase separation to obtain a pure product (ESI-MS: m/z=619.6 [M+H]+).
  • Figure US20230148436A2-20230511-C00147
  • Equimoles of EDCI and HOBt were added to the anhydrous DMF solution of SZU-101. After stirring for 15 minutes, DIPEA was added at 0° C. The resulting solution was reacted at room temperature for 0.5 hour, and then an equimole of Niraparib was added at 0° C. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water and filtered off with suction to give a crude product, which was purified by high-performance liquid chromatograph (HPLC) to obtain a pure product SZU-147 (ESI-MS: m/z=747.77[M+H]+).
  • Figure US20230148436A2-20230511-C00148
  • Lithium hydroxide and SZU-101 (in 1:1 ratio) were added to water. The resulting solution was stirred for 2-3 hours, and then lyophilized to remove the solvent to obtain a pure product.
  • Figure US20230148436A2-20230511-C00149
    Figure US20230148436A2-20230511-C00150
  • Synthesis of SZU-151:
  • After sodium salt was formed by adding metal Na to N, N-dimethylethanolamine, the starting materials were added to the reaction solution, heated to 95° C. and reacted for 3 hours. The solvent was removed under reduced pressure, and then water was added. The reaction solution was filtered off with suction to give a solid, which was washed with water to have a pH of 7-8 and dried under reduced pressure to obtain a pure product SZU-151.
  • Synthesis of SZU-152:
  • Bromine was added dropwise slowly to the acetic acid solution of SZU-151. After stirring overnight, the reaction solution was neutralized with NaHCO3. Small amount of ethanol was added into the obtained viscous compound. Subsequently, sodium thiosulfate solution was added to remove the remaining bromine. The reaction solution was filtered off with suction and dried to give a solid, and then 10% NaOH solution was added. The resulting solution was further filtered off with suction and dried under reduced pressure. The residue was purified by preparative liquid chromatography to obtain a bromide.
  • Metal Na was added to methanol. After stirring, the starting materials were added. The mixture was heated to 80° C. and reacted for 2 hours. Methanol was removed under reduced pressure. The resulted mixture was neutralized with HCl, filtered off with suction and dried under reduced pressure to give a solid SZU-152.
  • Synthesis of SZU-150 and SZU-153:
  • SZU-152 was added to HCl and stirred for 12 h. The solvent was removed under reduced pressure. The residue was separated by preparative liquid chromatography to give a compound SZU-150 (ESI-MS: 354.5 [M+H]+) and SZU-153 (ESI-MS: m/z=372.4 [M+H]+).
  • Figure US20230148436A2-20230511-C00151
  • NaOH was added to the ethanol/water (1:1) mixed solution of SZU-152 and refluxed overnight. The resulting solution was concentrated under reduced pressure, adjusted to have a pH of 3, and purified by preparative liquid chromatography to give a pure product SZU-154 (ESI-MS: m/z=387.5 [M+H]+).
  • Figure US20230148436A2-20230511-C00152
  • SZU-154 was dissolved in concentrated HCl and stirred overnight. The solvent was removed under reduced pressure. The residue was purified by preparative liquid chromatography to give a pure product SZU-155 (ESI-MS: m/z=373.5 [M+H]+).
  • Figure US20230148436A2-20230511-C00153
  • 50 mg of SZU-102 was dissolved in 0.5 mL of DMSO. 41 mg of tert-butoxycarbonyl (Boc)-protected lysine was added. The mixture was stirred at room temperature overnight. After the reaction was completed, DMSO was removed by lyophilization. The residue was purified by preparative liquid chromatography to give 69 mg of a white solid with a yield of 85%. ESI-MS: m/z=633.7 [M+H]+.
  • Figure US20230148436A2-20230511-C00154
  • 50 mg of SZU-102 was dissolved in 0.5 mL of DMSO. 47 mg of Boc-protected lysine was added. The resulting mixture was stirred at room temperature overnight. After the reaction was completed, DMSO was removed by lyophilization. The residue was purified by preparative liquid chromatography to give 68 mg of white solid with a yield of 83%. ESI-MS: m/z=533.6 [M+H]+.
  • Figure US20230148436A2-20230511-C00155
  • 109 mg of SZU-102 was dissolved in 1 mL of DMSO. 50 mg of PEG2 was added. The resulting mixture was stirred at room temperature overnight. After the reaction was completed, DMSO was removed by lyophilization. The residue was purified by preparative liquid chromatography to give 125 mg of a white solid with a yield of 79%. ESI-MS: m/z=564.6 [M+H]+.
  • Figure US20230148436A2-20230511-C00156
  • 87 mg of SZU-102 was dissolved in 0.9 mL of DMSO. 50 mg of PEG3 was added. The mixture was stirred at room temperature overnight. After the reaction was completed, DMSO was removed by lyophilization. The residue was purified by preparative liquid chromatography to give 105 mg of a white solid with a yield of 77%. ESI-MS: m/z=608.7 [M+H]+.
  • Figure US20230148436A2-20230511-C00157
  • 193 mg of a carboxylic compound was dissolved in 2 mL of DMSO. 100 mg of EDCI and 59 mg of N-hydroxysuccinimide (NHS) were added successively. After stirring at room temperature for 2 hours, the reaction solution was added with 50 mg of PEG2 and stirred at room temperature overnight. After the reaction was completed, DMSO was removed by lyophilization. The residue was purified by preparative liquid chromatography to give 203 mg of white solid with a yield of 73%. ESI-MS: m/z=519.5 [M+H]+.
  • Figure US20230148436A2-20230511-C00158
  • 155 mg of a carboxylic compound was dissolved in 1.5 mL of DMSO. 79 mg of EDCI and 47 mg of NHS were added successively. After stirring at room temperature for 2 hours, the reaction solution was added with 50 mg of PEG3 and stirred at room temperature overnight. After the reaction was completed, DMSO was removed by lyophilization. The residue was purified by preparative liquid chromatography to give 174 mg of a white solid with a yield of 72%. ESI-MS: m/z=563.6 [M+H]+.
  • Figure US20230148436A2-20230511-C00159
  • 0.5 g of metal Na was added slowly to 100 mL of anhydrous n-butanol at 0° C. After 30 minutes, 3 g of reactant a was added. The resulting mixture was stirred at 100° C. for 8 hours, cooled to room temperature and poured into a large amount of water. A yellow solid was precipitated out, filtered, washed with tetrahydrofuran three times, and dried to obtain 2.5 g of product b.
  • 2.5 g of the product b was mixed with 100 mL of chloroform and stirred homogeneously. 10 mL of bromine was added. The resulting solution was reacted at room temperature for 24 hours and filtered to obtain 2.2 g of product c, which was mixed with 60 mL of dioxane. Then 10 mL of concentrated hydrochloric acid was added. The mixture was reacted at 110° C. for 12 hours. The reaction mixture was poured into 600 mL of water and a crude solid product was precipitated out, which was filtered, dissolved in 60 mL of 1N NaOH, cooled and neutralized to a pH of 4 with hydrochloric acid. A pure product was precipitated out and then dried to give 1 g of SZU-149 with a yield of 55%. MS (ESI): m/z: [M+Na] 381.42.
  • 0.5 g of SZU-149 was dissolved in anhydrous methanol. 0.5 g of Raney Nickel was added. The resulting solution was subjected to hydrogenation reaction for 24 hours under 3 atmospheric pressures in hydrogen, filtered, and distilled under reduced pressure to give 0.4 g of a pure product SZU-163 with a yield of 89%; MS (ESI): m/z: [M+H] 329.41.
  • Figure US20230148436A2-20230511-C00160
  • Synthesis of SZU-166:
  • 1.5 g of SZU-163 was dissolved in 30 mL of DMSO. 1 mL of triethylamine and 0.5 g of ethyl chloroformate were added. The mixture was stirred at room temperature for 2 hours. The reaction mixture was distilled under vacuum to a small volume, cooled to room temperature, and added with 30 mL of ethyl acetate. A solid was precipitated out, filtered, washed with ethyl acetate, and dried to give 0.75 g of a pure product SZU-166 with a yield of 46%; MS (ESI): m/z: [M+H]355.40.
  • Synthesis of SZU-181 (Conjugate Formed by Conjugating SZU-166 with Hepatitis B PreS Antigen Epitope):
  • Figure US20230148436A2-20230511-C00161
  • A mixture of SZU-166 (10 mg) and PreS-peptide (90 mg) was dissolved in 3 mL of DMF. 0.2 mL of triethylamine was added. The mixture was reacted under stirring at 20° C. for 24 hours. 3 mL of dichloromethane was added. The precipitated product was separated by centrifugation to give a crude product, which was purified by HPLC, and lyophilized to give 65 mg of SZU-181. Molecular weight determined by mass spectrometry: 3512.80: (1756.3×2), (1170.9×3).
  • Figure US20230148436A2-20230511-C00162
    Figure US20230148436A2-20230511-C00163
  • EDCI and HOBt were added to an equimole of 4-pentynoic acid in DMF. After 15 minutes, triethylamine was added. The mixture was reacted under stirring at 0° C. for 15 minutes, and then the reaction was continued under stirring at room temperature for 30 minutes. Subsequently, an equimole of TL-008 was added. The resulting mixture was reacted at room temperature overnight. The reaction solution was poured into water. The precipitated product was filtered and dried under vacuum to give SZU-136 with a yield of 87%; MS (ESI): m/z: [M+1] 425.51.
  • SZU-136, azide compound 2 and sodium ascorbate in equimolar amounts were added to the solution of DMSO and water (1:1). The resulting solution was stirred at room temperature overnight, added with 10-fold volume of water and filtered. The obtained product was purified by HPLC, and dried to obtain SZU-139 with a yield of 77%; MS (ESI): m/z: [M+1] 654.72.
  • Figure US20230148436A2-20230511-C00164
  • SZU-134 was dissolved in DMSO. Equimoles of NHS and EDCI were added. After stirring for 10 minutes, an equimole of Direct violet N (OX40 agonist) was added. The reactants were reacted in a closed vessel at room temperature for 3 days. 20-fold amount of water was added. Solid was precipitated out and filtered, and then 2N NaOH was added. The mixture was stirred at 10° C. for 30 minutes and filtered. The filtrate was acidified with 2N hydrochloric acid to have a pH of 1 and gave a precipitated solid product SZU-169 with a yield of 52%; MS (ESI): m/z: [M+2Na] 1611.64.
  • Figure US20230148436A2-20230511-C00165
    Figure US20230148436A2-20230511-C00166
  • The reactant a was dissolved in small amount of DMF. One-fold equivalent amount of sodium azide was added. The mixture was reacted under stirring at room temperature for 24 hours. The reaction mixture was added with 10-fold volume of water and extracted with ethyl acetate. The extracted phase was washed with water three times. The solvent was distilled off under reduced pressure to obtain product b with a yield of 90%.
  • 100 mg of Afatinib and 50 mg of the product b were dissolved in 15 mL of DMF. Trace amount of NaI was added. The mixture was reacted at 60° C. for 24 hours. 100 mL of n-butanol was added to the reaction mixture, which was subjected to distillation under reduced pressure to a small volume (about 5 mL), cooled to room temperature, and added with 50 mL of ethyl acetate. A solid product c was precipitated out and dried under vacuum to give 118 mg of product c; MS (ESI): m/z: [M+H] 688.21.
  • 100 mg of the product c, 60 mg SZU-136, 10 mg of copper sulfate, and 30 mg of vitamin C were added to the solution of DMSO and water (1:1). The resulting solution was stirred at room temperature overnight, added with 10-fold volume of water and filtered. The obtained product was purified by HPLC, and dried to obtain 105 mg of SZU-174 with a yield of 65%; MS (ESI): m/z: [M+1] 1112.70.
  • Figure US20230148436A2-20230511-C00167
  • 5 mg of SZU-158 was dissolved in 0.5 mL of DMSO. Equimoles of NHS and EDCI were added. The mixture was stirred at room temperature for 4 hours. 20 mg of PD-1 antibody (purchased from Bioxcell, J43) in 0.5 mL of PBS was added. The mixture was stirred in a closed vessel at 15° C. overnight. The mixed reaction solution was filtered with a filter membrane to obtain a SZU-158-PD-1 conjugate product, which was lyophilized to obtain 18 mg of a white solid SZU-158-PD-1. The conjugation degree determined by mass spectrometry is n=4 (SZU-158-PD-1 has immunocytokine activities as shown in FIGS. 58 and 65 , and antibody functions as shown in FIGS. 58, 64 and 65 ).
  • Figure US20230148436A2-20230511-C00168
  • 5 mg of SZU-158 was dissolved in 0.5 mL of DMSO. Equimoles of NHS and EDCI were added. The mixture was stirred at room temperature for 4 hours. 20 mg of OX40 antibody (purchased from Bioxcell, OX-86) in 0.5 mL of PBS was added. The resulting mixture was stirred in a closed vessel at 15° C. overnight. The mixed reaction solution was filtered with a filter membrane to obtain SZU-158-OX40 conjugate product, which was lyophilized to obtain 17 mg of a white solid SZU-158-OX40. The conjugation degree determined by mass spectrometry is n=4 (SZU-158-OX40 has immunocytokine activities as shown in FIGS. 58 and 65 , and antibody functions as shown in FIGS. 58, 64 and 65 ).
  • Figure US20230148436A2-20230511-C00169
  • 5 mg of SZU-158 was dissolved in 0.5 mL of DMSO. Equimoles of NHS and EDCI were added. The mixture was stirred at room temperature for 4 hours. 20 mg of PD-L1 antibody (purchased from Bioxcell, 1° F.9G2) in 0.5 mL of PBS was added. The mixture was stirred in a closed vessel at 15° C. overnight. The mixed reaction solution was filtered with a filter membrane to obtain SZU-158-PD-L1 conjugate product, which was lyophilized to obtain 15 mg of a white solid SZU-158-PD-L1. The conjugation degree determined by mass spectrometry is n=4 (SZU-158-PD-L1 has immunocytokine activities as shown in FIGS. 58 and 65 , and antibody functions as shown in FIGS. 58, 64 and 65 ).
  • The novel immune-targeted compounds formed by conjugating SZU-158 with an antibody, owing to the presence of dimethylamino group, can form a salt and result in an ionized positive charge, which is advantageous for improving the therapeutic efficacy and water solubility.
  • SZU-168 and SZU-175
  • Vemu-1, a precursor of Vemurafenib, is purchased from WuXi AppTec.
  • Figure US20230148436A2-20230511-C00170
    Figure US20230148436A2-20230511-C00171
    Figure US20230148436A2-20230511-C00172
  • Vemu-1 was dissolved in DMF. 2-fold moles of triethylamine were added. The mixture was cooled to 0° C. and then added with 1-fold mole of vinylsulfonyl chloride. The resulting mixture was warmed slowly to room temperature, stirred for 8 hours, and then added with 10-fold volume of ice-water. The mixture was stirred thoroughly for 1 hour and adjusted with saturated Na2CO3 to have a pH of 8. The precipitated solid was filtered, washed with water twice, and dried to give Vemu-2. 1 g of Vemu-2 was dissolved in 30 mL of DMF. 0.33 g of mercaptoethylamine was added. The mixture was stirred at room temperature for 12 hours. The reaction mixture was distilled off the solvent under reduced pressure to obtain crude Vemu-3, which was purified by silica gel column chromatography to obtain 0.98 g of a pure product Vemu-3 with a yield of 85%; MS (ESI): m/z: [M+1] 552.04.
  • 0.3 g of Vemu-3 was dissolved in 10 mL of DMSO. 0.2 g of SZU-166 was added. The mixture was reacted under stirring at 40° C. for 8 hours, and then added with 100 mL of water. The reaction solution was centrifuged to give precipitated crude product, which was dissolved in 20 mL of glacial acetic acid. After filtration, the filtrate was added with 100 mL of water and a pure product SZU-168 was precipitated, which was dried to 0.33 g; MS (ESI): m/z: [M+1]906.41.
  • 1 g of SZU-158 was dissolved in 10 mL of DMSO. 0.2 g of NHS (N-hydroxysuccinimide) and 0.33 g of EDCI (1-(3-dimethylaminopropyl)-3-ethylcarbodiimide hydrochloride) were added. The mixture was left under stirring at room temperature overnight to obtain SZU-137. 0.9 g of Vemu-3 was added and stirring was continued at room temperature for another 12 hours. The mixture was added with 100 mL of water and filtered to give a crude product, which was re-crystallized with acetic acid to obtain 1.22 g of SZU-175 acetate; MS(ESI): m/z: [M+1]1095.81.
  • SZU-176
  • Intermediate of Osimertinib was purchased from WuXi AppTec.
  • Figure US20230148436A2-20230511-C00173
  • 1 g of SZU-144 was dissolved in 10 mL of DMSO. 0.26 g of NHS and 0.4 g of EDCI were added. The mixture was left under stirring at room temperature overnight. 1 g of intermediate of Osimertinib was added, and then the stirring was continued at room temperature for another 12 hours. The mixture was added with 100 mL of water and filtered to give a crude product, which was re-crystallized with acetic acid to obtain 1.5 g of SZU-176 acetate; MS (ESI): m/z: [M+1] 870.81.
  • Figure US20230148436A2-20230511-C00174
  • 100 mg of Erlotinib, 150 mg of SZU-160, 20 mg of copper sulfate and 20 mg of vitamin C were added to a solution (10 mL) of DMSO and water (1:1). The mixture was left under stirring at room temperature overnight. 10-fold volume of water was added. The reaction solution was filtered, purified by HPLC and dried to give 176 mg of SZU-177 with a yield of 70%; MS (ESI); m/z: [M+1] 993.10.
  • Figure US20230148436A2-20230511-C00175
  • 3.5 mg of SZU-160 was dissolved in 0.5 mL of DMSO. Equimoles of NHS and EDCI were added. The mixture was left under stirring at room temperature for 4 hours. 20 mg of PD-L1 antibody (purchased from Bioxcell, 10F.9G2) in 0.5 mL of PBS was added. The mixture was stirred in a closed vessel at 15° C. overnight. The mixed reaction solution was filtered with a filter membrane to obtain a solution of SZU-160-PD-L1 antibody conjugate product. 2.5 mg of Erlotinib, 2 mg of sodium ascorbate, and 2 mg of copper sulfate were added. The mixture was reacted under stirring at room temperature for 12 hours. The reaction solution was filtered with filter membrane and lyophilized to give 12 mg of a white solid SZU-178 (i.e., SZU-160-Erlotinib-PD-L1 conjugate formed by conjugating SZU-178 with Erlotinib and PD-L1 antibody). The conjugation degree determined by mass spectrometry is n=4 (SZU-178 has immune cytokine stimulation and anti-tumor functions as shown in FIGS. 58, 64 and 65 ).
  • Figure US20230148436A2-20230511-C00176
  • 71 mg of miRNA21 (purchased from Suzhou Jima Gene Co., Ltd.) was dissolved in 10 mL of acetonitrile and water (1:1) solvent. 14 mg of Ag2CO3 was added. The mixture was stirred at room temperature for 12 hours. The solvent was distilled off under reduced pressure to give 69 mg of Ag-miRNA21 (96%), which was mixed with 5 mg of chlorinated azide in 20 mL of acetonitrile. The mixture was heated under reflux for 4 hours, cooled to room temperature, and added with 50 mL of water. The resulting mixture was stirred for 15 minutes and filtered. The filtrate was distilled under reduced pressure to a small volume (10 mL). 50 mL of tetrahydrofuran was added. After stirring for 10 minutes, the solution was centrifuged to give precipitate, which was dried under vacuum to obtain 45 mg of N3-miRNA21 (yield 61%). MS (ESI): m/z: [M+Na] 7307.42.
  • 40 mg of N3-miRNA21, 3 mg of SZU-136, 2 mg of copper sulfate and 2 mg of vitamin C were mixed and added to a solution (10 mL) of DMSO and water (1:1). The mixture was stirred at room temperature overnight. The product was purified by HPLC, lyophilized, and dried under vacuum to give 14 mg of SZU-136-miRNA21 (i.e., the conjugate formed by conjugating SZU-136 with miRNA21) with a yield of 33%; MS (ESI): m/z: [M+Na] 7731.86.
  • Figure US20230148436A2-20230511-C00177
  • 580 mg of SZU-115, 267 mg of Zidovudine, 25 mg of copper sulfate and 25 mg of sodium ascorbate were admixed and added to a solution (25 mL) of DMSO and water (1:1). The mixture was stirred at room temperature overnight and added with 10-fold volume of water. The reaction solution was distilled under vacuum to a small volume (about 10 mL) and added with 30 mL of water. The mixed solution was adjusted to a pH of 10 with 5% NaOH and filtered. The filtrate was adjusted to a pH of 5 with glacial acetic acid and gave precipitated product, which was filtered and dried under vacuum to obtain 728 mg of SZU-179 with a yield of 86%; MS (ESI): m/z: [M+Na] 871.42.
  • SZU-115-PD-L1-targeted drug (i.e., the conjugate formed by conjugating SZU-115 with PD-L1 antibody and targeted drug)
  • Figure US20230148436A2-20230511-C00178
  • Conjugate of SZU-115-antibody-targeted drug
  • Figure US20230148436A2-20230511-C00179
  • The PD-L1 antibody was conjugated by the same process as in the preparation of SZU-178, and Click reaction was conducted by the same process as in the preparation of SZU-179, to obtain SZU-180. The resulting product can be used for the treatment of AIDS.
    • EFFECT EXAMPLES Effect Example 1: Detection of the Immunostimulatory Effects of the SZU-Series Small-Molecule Immune Agonists and Novel Immune Targeted Compounds of the Present Invention on Spleen Lymphocytes
  • 1. Extraction of Spleen Lymphocytes
  • (1) BALB/c or C57BL/6 mouse spleens were collected, ground in a small dish containing Mouse Lymphocyte Separation Medium, and filtered through a filter membrane (with a pore size of 0.22 μm) to obtain a cell suspension, which was treated by centrifugation to obtain lymphocytes. The isolated lymphocytes were placed in a 15 mL centrifuge tube, into which 10 mL of 1640 complete medium was added. The mixture was mixed uniformly and centrifuged at 250 g for 10 minutes. The supernatant was discarded.
  • (2) The cells were re-suspended with 1 mL of erythrocyte lysate. After 1 minute of lysis, 4 mL of 1640 complete medium was added for termination. The tube was centrifuged at 1,000 rpm for 5 minutes. The supernatant was discarded.
  • (3) The cells were re-suspended with 2 mL of 1640 complete medium, counted, and then seeded into 24-well plates at a density of 1×106 cells per well.
  • 2. Preparation of the Samples
  • The spleen lymphocytes seeded into the 24-well plates were stimulated with different concentrations of the SZU-series small-molecule immune agonists and novel immune targeted compounds. The 24-well plate was transferred into an incubator. After 24 hours of stimulation, the supernatant was gently aspirated, i.e. samples to be tested by ELISA (Enzyme-Linked Immunosorbent Assay). The concentrations and concentration gradients were dependent on individual requirements of each tested sample.
  • 3. ELISA Test
  • (1) Coating: 10× coating buffer was diluted to 1× and then used to dilute the capture antibody. The diluted capture antibody was added into a 96-well microtiter plate, 100 μl per well. The plate was sealed with plastic wrap and placed at 37° C. for 2 to 4 hours or at 4° C. in refrigerator overnight. The liquid contents were aspirated out of each well of the plate. Then the plate was washed with washing buffer (PBST:PBS solution with 5% Tween-20) three times and dried by patting.
  • (2) Blocking: diluted blocking solution was added, 200 μl per well; the plate was sealed with plastic wrap and blocked at room temperature for 1 hour. Then the plate was washed with PBST three times and dried by patting.
  • (3) Loading samples: 12 standard samples were arranged, wells in duplicate for each concentration of a standard sample, with the highest concentration of the standard sample being 2,000 pg/mL. The samples were diluted with dilution buffer to the desired concentration. The standard samples have the following successive concentrations: 2,000 pg/mL, 1,000 pg/mL, 500 pg/mL, 250 pg/mL, 125 pg/mL, 62.5 pg/mL, 31.5 pg/mL, 15.75 pg/mL, 7.875 pg/mL, 3.9375 pg/mL, 1.96875 pg/mL, and 0 pg/mL. The supernatant samples obtained by stimulating spleen lymphocytes with different concentrations of drugs were added into the corresponding wells, wells in triplicate per sample. Blank control wells and negative control wells were also arranged and labeled accordingly. The 96-well microtiter plate loaded with the standard samples and tested samples was sealed with plastic wrap and incubated at room temperature for 2 hours or at 4° C. in refrigerator overnight. The liquid contents were aspirated out of each well. The plate was washed with PBST three times and dried by patting.
  • (4) Secondary antibody: the testing antibody was diluted with dilution buffer to the desired concentration, and added to the plate at 100 μl per well, and incubated at room temperature for 1 hour. The liquid contents were aspirated out of each well of the plate. Then the plate was washed with PBST five times and dried by patting.
  • (5) Enzyme-labeled antibodies: Avidin-HRP was diluted with dilution buffer to the desired concentration, and added to the plate at 100 μl per well, and then incubated at room temperature for 30 minutes. The liquid contents were aspirated out of each well of the plate. Then the plate was washed with PBST five times and dried by patting.
  • (6) Color development: 100 μl of 3,3′,5,5′-tetramethylbenzidine (TMB) chromogenic solution was added to each well, and then incubated at room temperature in the dark for 15 minutes.
  • (7) Termination: after the completion of color development, 50p of stop solution was added to each well, wherein the stop solution was 1 mol/L H2SO4.
  • (8) Detection: the 96-well microtiter plate after termination was placed into a full-wavelength microplate reader, and the absorbance values were read at the wavelength of 450 nm. The data were exported and plotted as a standard curve with software; and a linear regression equation was derived. The concentrations of the immune factors (INF-γ, IL-6, etc.) produced by immunocytes were calculated. Specific results of the experiments are given in the figures attached hereto.
    • Effect Example 2: Anti-Tumor Effects of the Novel Immune Targeted Compounds of the Present Invention
  • 1. Tumor Treatment Protocol in Mice
  • (1) BALB/c or C57BL/6 mice of 6-8 weeks old were weighed and anesthetized with pentobarbital sodium (50 mg/kg i.p). Hair was removed from the tumor implantation site (on the back). The mice were immobilized with sticky tapes.
  • (2) Surgical sites were sterilized by using cotton swabs dipped into 75% alcohol. Small incisions of about 1 cm at the tumor implantation sites were made and injected with 100 μl of cancer cell suspension (5×104 CT26 cells, B16 cells or 4T1 cells). The incisions were sealed with surgical thread and sterilized. After the operations, 50,000 units of ampicillin/100 μL/mouse were injected intraperitoneally for 3 consecutive days.
  • 2. Administration
  • The mice were administered with PBS blank control, the immune agonists or novel immune targeted compounds, respectively. The dosages were determined according to the actual needs (following the principle of effective and safe dosages). The respective substance was dissolved in an appropriate solvent, and the volume of each administration was 100 μL; the mice in each group were administered via intraperitoneal injection. Each group was administered on day 7, 15, 22, and 29 after tumor implantation; and the tumor sizes were measured regularly. The mice were euthanized when the tumor size reached 1500 mm3 or had a weight higher than 15% of their body weights. The results of tumor suppression and survival rates were shown in FIGS. 57, 58 and 64 . It can be seen that the groups of mice treated with the novel immune targeted compounds according to the present invention have significantly higher anti-tumor effect.
  • Methods for testing the biological activities of the novel small-molecule immune agonists and novel immune targeted compounds of the present invention are not limited to the above test methods, and other known or accepted activity testing methods may also be used. Analogously, the schematic diagrams of activities in the present invention are not limited to the illustrated schematic diagrams, while these schematic diagrams can inspire those skilled in the art to achieve the application effects of the novel small-molecule immune agonists and novel immune targeted compounds according to the present invention.

Claims (15)

1. Small-molecule immune agonist of Formula (I), or isomer or pharmaceutically acceptable salt thereof:
Figure US20230148436A2-20230511-C00180
wherein
A1 represents O or C;
A2 represents a nitro, amino, isocyanate, alkyleneamino, alkylene isocyanate, or a group selected from Formula (II) or (III):
Figure US20230148436A2-20230511-C00181
wherein
R1 and R2 each independently represent (CH2)p(NH)q, where p is 0, 1 or 2, and q is 0, 1 or 2;
R3 represents O or S;
R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0, 1 or 2; z is 0, 1 or 2; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0, 1 or 2, wherein when s≠0 and t=0, the carbon atom adjacent to the carbonyl group in (CH2)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
Figure US20230148436A2-20230511-C00182
or forming a double bond with another carbon atom;
R5 represents hydroxy, alkoxy, hydroxylamine, vinyl group or a group selected from:
Figure US20230148436A2-20230511-C00183
R6 represents alkyl or a group selected from:
Figure US20230148436A2-20230511-C00184
where * denotes the site linked to the carbon atom;
with the proviso that the following compounds are excluded:
Figure US20230148436A2-20230511-C00185
Figure US20230148436A2-20230511-C00186
2. The small-molecule immune agonist of Formula (I), or isomer or pharmaceutically acceptable salt thereof according to claim 1,
Figure US20230148436A2-20230511-C00187
wherein
A1 represents O or C;
A2 represents nitro, amino, isocyanate, C1-C6 alkylene amino, C1-C6 alkylene isocyanate or a group selected from formula (II) or (III):
Figure US20230148436A2-20230511-C00188
wherein
R1 and R2 each independently represent (CH2)p(NH)q, where p is 0 or 1; and q is 0 or 1;
R3 represents O or S;
R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0 or 1; z is 0 or 1; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0 or 1, wherein when s≠0 and t=0, the carbon atom adjacent to the carbonyl group in (CH2)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
Figure US20230148436A2-20230511-C00189
or forming a double bond with another carbon atom;
R5 represents hydroxyl, C1-C6 alkoxy, hydroxylamine, vinyl group or a group selected from:
Figure US20230148436A2-20230511-C00190
R6 represents C1-C6 alkyl group or a group selected from:
Figure US20230148436A2-20230511-C00191
wherein * denotes the site linked to the carbon atom;
with the proviso that the following compounds are excluded:
Figure US20230148436A2-20230511-C00192
3. The small-molecule immune agonist of Formula (I), or isomer or pharmaceutically acceptable salt thereof according to claim 2,
Figure US20230148436A2-20230511-C00193
wherein,
A1 represents O or C;
A2 represents nitro, amino, isocyanate, methylene amino, methylene isocyanate or a group selected from Formula (II) or Formula (III):
Figure US20230148436A2-20230511-C00194
wherein,
R1 and R2 each independently represent (CH2)p(NH)q, where p is 0 or 1, and q is 0 or 1;
R3 represents O or S;
R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0 or 1; z is 0 or 1; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0 or 1, wherein when s≠0 and t=0, the carbon atom adjacent to the carbonyl group in (CH9)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
Figure US20230148436A2-20230511-C00195
or forming a double bond with another carbon atom;
R5 represents a hydroxyl, methoxy, hydroxylamine, vinyl group or a group selected from:
Figure US20230148436A2-20230511-C00196
R6 represents a methyl group or a group selected from:
Figure US20230148436A2-20230511-C00197
wherein * denotes the site linked to the carbon atom;
with the proviso that the following compounds are excluded:
Figure US20230148436A2-20230511-C00198
4. The small-molecule immune agonist of Formula (I), or isomer or pharmaceutically acceptable salt thereof according to claim 3, which is a SZU-series compound selected from the group consisting of: SZU-104, SZU-105, SZU-107, SZU-108, SZU-109, SZU-110, SZU-111, SZU-112, SZU-113, SZU-115, SZU-118, SZU-120, SZU-127, SZU-128, SZU-129, SZU-131, SZU-132, SZU-133, SZU-134, SZU-135, SZU-136, SZU-137, SZU-138, SZU-139, SZU-140, SZU-142, SZU-143, SZU-144, SZU-145, SZU-149, SZU-158, SZU-159, SZU-160, SZU-161, SZU-162, SZU-163, SZU-166 and SZU-171, or pharmaceutically acceptable salt thereof.
5. Immune targeted compound or pharmaceutically acceptable salt thereof, which is formed by covalently conjugating a small-molecule immune agonist or an addition product formed from the immune agonist and a mercapto-containing polypeptide such as glutathione, with targeted drug or antibody, and represented by the following general formula:

(TLA)n-L-(Tar)m
wherein TLA represents small-molecule immune agonist or its addition product with mercapto-containing polypeptide such as glutathione; Tar represents targeted drug or antibody; n and m each independently represent a number ranging from 1 to 5; L represents a linker, which is linear or branched alkane or polyethylene glycol chain, and both ends of L are covalently linked to TLA and/or Tar directly or via additional groups.
6. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, wherein the small-molecule immune agonist comprise the small-molecule immune agonist of Formula (I), or isomer or pharmaceutically acceptable salt thereof:
Figure US20230148436A2-20230511-C00199
wherein
A1 represents O or C;
A2 represents a nitro, amino, isocyanate, alkyleneamino, alkylene isocyanate, or a group selected from Formula (II) or (III):
Figure US20230148436A2-20230511-C00200
wherein
R1 and R2 each independently represent (CH2)p(NH)q, where p is 0, 1 or 2, and q is 0, 1 or 2;
R3 represents O or S;
R4 represents (CH2)x(C═O)y(NH)z[(CH2)2O]r(CH2)s(C═C)t, where x is an integer ranging from 0 to 5; y is 0, 1 or 2; z is 0, 1 or 2; r is 0, 1, 2 or 3; s is an integer ranging from 0 to 5 and t is 0, 1 or 2, wherein when s≤0 and t=0, the carbon atom adjacent to the carbonyl group in (CH2)s may be substituted by a group selected from —NHBoc, an amino group, —NHFmoc,
Figure US20230148436A2-20230511-C00201
or forming a double bond with another carbon atom;
R5 represents hydroxy, alkoxy, hydroxylamine, vinyl group or a group selected from:
Figure US20230148436A2-20230511-C00202
R6 represents alkyl or a group selected from:
Figure US20230148436A2-20230511-C00203
where * denotes the site linked to the carbon atom.
7. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, wherein the small-molecule immune agonist is a SZU-series compound selected from the group consisting of: SZU-102, SZU-104, SZU-105, SZU-106, SZU-107, SZU-108, SZU-109, SZU-110, SZU-111, SZU-112, SZU-113, SZU-115, SZU-118, SZU-120, SZU-127, SZU-128, SZU-129, SZU-131, SZU-132, SZU-133, SZU-134, SZU-135, SZU-136, SZU-137, SZU-138, SZU-139, SZU-140, SZU-142, SZU-143, SZU-144, SZU-145, SZU-149, SZU-158, SZU-159, SZU-160, SZU-161, SZU-162, SZU-163, SZU-166, SZU-171, SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 and SZU-130, or pharmaceutically acceptable salt thereof.
8. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, wherein the TLA represents a compound selected from the group consisting of: SZU-103-GSH, SZU-114-GSH, SZU-117-GSH, SZU-122-GSH, SZU-127-GSH, SZU-133-GSH, SZU-138-GSH and SZU-144-GSH.
9. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, wherein the Tar represents at least one of the compounds selected from the group consisting of: Dasatinib, Imatinib, Saracatinib, Ponatinib, Nilotinib, Danusertib, AT9283Degrasyn, Bafetinib, kw-2449, nyp-bhg712, dcc-2036, GZD824, GNF-2, PD173955, GNF-5, Bosutinib, Gefitinib, Erlotinib, Sunitinib, Ruxolitinib, Tofacitinib, Lapatinib, Vandetanib, Sorafenib, Sunitinib, Axitinib, Nintedanib, Regorafenib, Pazopanib, Lenvatinib, Crizotinib, Ceritinib, Cabozantinib, DWF, Afatinib, Ibrutinib, Niraparib, Palbociclib, B43, KU004, Foreinib, KRCA-0008, PF-06439015, PF-06463922, Canerlinib, GSA-10, GW2974, GW583340, W24002, CP-380736, D2667, Mubritinib, PD153035, PD168393, Pelitinib, PF-06459988, PF06672131, PF-6422899, PK1166, ReveromycinA, Tyrphostin 1, tyrphostin23, lyrphostin51, Tyrphostin, AG528, Tyrphostin, AG658, Tyrphostin, AG825, Tyrphostin, AG835, Tyrphostin, AG1478, Tyrphostin, RG13022, Tyrphostin, RG14620, B178, GSK1838705A, PD-161570, PD173074, SU-5402, Roslin2, Picropodophyllotoxin, PQ401, i-ome-tyrphostin, AG538, GNF5837, GW441756, Tyrphostin, AG879, DMPQ, jnj-10198409, PLX647, Trapidil, TyrphostinA9, Tyrhostin, AG370, Lestaurtinib, DMH4, Eldanamvcin, Genistein, Gw2580, HerbimycinA, Lavendustin, CMidostaurin, nvp-bhg712, PD158780, pd-166866, pf-0627334, PP2, RPL, SUl1274, SU5614, Symadex, Tyrphostin, AG34, Tyrphostin, AG974, Tyrphostin, AG1007, UNC2881, Honokiol, SU1498, SKLB1002, cp-547632, jk-p3, KRN633, sc-1, ST638, SU 5416, Sulochrin, Tyrphostin, SU1498, rociletinib, Dacomitinib, Tivantinib, Neratinib, Masitinib, Vatalanib, Icotinib, xl-184, osi-930, AB1010, Quizartinib, AZD9291, Tandutinib, HM61713, Brigantinib, Vemurafenib (plx-4032), Semaxanib, AZD2171, Crenolanib, Damnacanthal, Fostamatinib, Motesanib, Radotinib, osi-027, Linsitinib, BIX02189, PF-431396, PND-1186, PF-03814735, PF-431396, sirollmus, temsirolimus, everolimus, deforolimus, Zotarolimus, BEZ235, INK128, Omipalisib, AZD8055, MHY1485, PI-103, KU-0063794, ETP-46464, GDC0349, XL388, WYE-354, WYE-132, GiSK1059615, WAY-600, PF-04691502, wYE-687, PP121, BGT226, AZD2o014, PP242, CH5132799, P529, GDC0980, GDC-0994, XMD82, Ulixertinib, FR180204, SCH772984, Trametinib, PD184352, PD98059, Selumetinib, PD325901, U0126, Pimasertinib, tak-733, AZD8330, Binimetinib, PD318088, SL-327, Refametinib, gdc-0623, Cobimetinib, b1-847325, Adaphostin, GNF2, PPYA, aim-100, ASP 3026, LFM, Toceranib, JQ1, Niraparib, Fuzuopali, palbociclib, ARS-853, ARS-1620, Chlorazol-violet N (Direct violet N), miRNA-21, PreS, Zidovudin, Lenvatinib, LY-364947, ARS-1620 or Reparixin, EGFR inhibitors, TKIs, BRD4 inhibitors, KRAS pathway-related target inhibitors, BRAF inhibitors, BTK Inhibitors, PARP inhibitors, PD-L1 inhibitors (including antibodies and small-molecules), PD-1 inhibitors (including antibodies and small-molecules), OX40 agonists (including antibodies and small-molecules), CD122 antibodies, CD3 antibodies, CD19 inhibitors, CD20 inhibitors, MUC1 inhibitors, MUC16 inhibitors, CDK4/6 inhibitors, TGF-β inhibitors, CXCR inhibitors, CCL and CXCL chemokine inhibitors and miRNAs.
10. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 9, wherein the Tar represents at least one of the compounds selected from the group consisting of: TKIs, EGFR inhibitors, KRAS inhibitors, BRAF inhibitors, BTK inhibitors, BRD4 inhibitors, PD-L1 inhibitors, PD-1 inhibitors, OX40 agonists, PARP inhibitors and CDK4/6 inhibitors.
11. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, wherein L is
Figure US20230148436A2-20230511-C00204
12. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, wherein the additional groups include amides, alkoxy bonds, alkylthio bonds, substituted amine groups, quaternary amine salts, heterocycles formed by Click reaction, amino acid groups that are dissociable by proteases, ureido or thiourea groups.
13. The immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5, which is a SZU-series compounds selected from the group consisting of: SZU-116, SZU-119, SZU-124, SZU-125, SZU-146, SZU-147, SZU-168, SZU-169, SZU-174, SZU-175, SZU-176, SZU-177, SZU-178, SZU-179, SZU-180, SZU-181, SZU-158-PD-L1, SZU-158-OX40, SZU-158-PD-1 or SZU-136-miRNA21, or pharmaceutically acceptable salt thereof.
14. A method of immunomodulation comprising administering to a subject the small-molecule immune agonist of formula (I) or isomer or pharmaceutically acceptable salt thereof according to claim 1, compound SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 or SZU-130 or the immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5.
15. A method for treating a tumor, for defending against a virus, or for eliminating a target protein, comprising administering to a subject the small-molecule immune agonist of formula (I) or isomer or pharmaceutically acceptable salt thereof according claim 1, compound SZU-101, SZU-103, SZU-114, SZU-117, SZU-122 or SZU-130 or the immune targeted compound or pharmaceutically acceptable salt thereof according to claim 5.
US17/044,659 2018-04-03 2019-04-02 Small molecule immune agonists and immune targeting compounds and application thereof Active 2042-08-19 US12281117B2 (en)

Applications Claiming Priority (5)

Application Number Priority Date Filing Date Title
CN201810289351.7 2018-04-03
CN201810289351 2018-04-03
CN201810331582.X 2018-04-13
CN201810331582.XA CN108379591B (en) 2018-04-03 2018-04-13 Synthesis of immune agonist targeting compound and application thereof
PCT/CN2019/080945 WO2019192454A1 (en) 2018-04-03 2019-04-02 Novel small molecule immune agonists and immune targeting compounds and application thereof

Publications (3)

Publication Number Publication Date
US20210038605A1 US20210038605A1 (en) 2021-02-11
US20230148436A2 true US20230148436A2 (en) 2023-05-11
US12281117B2 US12281117B2 (en) 2025-04-22

Family

ID=63073860

Family Applications (1)

Application Number Title Priority Date Filing Date
US17/044,659 Active 2042-08-19 US12281117B2 (en) 2018-04-03 2019-04-02 Small molecule immune agonists and immune targeting compounds and application thereof

Country Status (4)

Country Link
US (1) US12281117B2 (en)
JP (1) JP2021519792A (en)
CN (2) CN115192726A (en)
WO (1) WO2019192454A1 (en)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12172992B2 (en) 2022-03-22 2024-12-24 Abbvie Inc. Pyrimidines for degrading Bruton's tyrosine kinase

Families Citing this family (31)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
IL263616B2 (en) 2016-07-07 2025-01-01 Univ Leland Stanford Junior Antibody adjuvant conjugates
CN107281483B (en) * 2017-04-25 2025-10-24 深圳大学 Immune agonist and composition and application thereof and preparation method of the composition
JP2021512105A (en) 2018-01-31 2021-05-13 デシフェラ・ファーマシューティカルズ，エルエルシー Combination therapy for the treatment of gastrointestinal stromal tumors
JP7523351B2 (en) 2018-01-31 2024-07-26 デシフェラ・ファーマシューティカルズ，エルエルシー Combination Therapy for the Treatment of Mastocytosis - Patent application
CN115192726A (en) 2018-04-03 2022-10-18 深圳大学 Synthesis of immune agonist targeting compound and application thereof
CN109796407A (en) * 2018-12-29 2019-05-24 鼎元(天津)生物医药科技有限公司 A kind of modularization prepares the method and application of chemical small molecule probe
AU2020241686A1 (en) 2019-03-15 2021-11-04 Bolt Biotherapeutics, Inc. Immunoconjugates targeting HER2
NZ784949A (en) 2019-08-12 2025-09-26 Deciphera Pharmaceuticals Llc Ripretinib for treating gastrointestinal stromal tumors
TWI878335B (en) 2019-08-12 2025-04-01 美商迪賽孚爾製藥有限公司 Methods of treating gastrointestinal stromal tumors
JP2023500385A (en) * 2019-11-11 2023-01-05 シーストーン・ファーマシューティカルズ・（スージョウ）・カンパニー・リミテッド Pharmaceutical combinations and uses thereof
AU2020419197B2 (en) 2019-12-30 2023-08-31 Deciphera Pharmaceuticals, Llc Amorphous kinase inhibitor formulations and methods of use thereof
NZ789199A (en) 2019-12-30 2025-07-25 Deciphera Pharmaceuticals Llc Compositions of 1-(4-bromo-5-(1-ethyl-7-(methylamino)-2-oxo-1,2-dihydro-1,6-naphthyridin-3-yl)-2-fluorophenyl)-3-phenylurea
CN111704614B (en) * 2020-04-15 2021-10-22 深圳大学 A series of immune agonists
CN111892648B (en) * 2020-06-08 2022-08-26 中国科学院上海药物研究所 Novel coronavirus polypeptide vaccine coupled with TLR7 agonist and application thereof
CN111643661A (en) * 2020-06-08 2020-09-11 王竹林 Novel immunologic adjuvant, compound and application thereof
CN111704603B (en) * 2020-06-12 2021-10-26 四川省人民医院 Anti-tumor compound and application thereof
CN112336852A (en) * 2020-10-14 2021-02-09 深圳大学 Whole cell vaccine and preparation method and application thereof
CN114533879B (en) * 2020-11-19 2023-09-29 广州顺健生物医药科技有限公司 Combination therapy for the treatment of cancer
CN112457297B (en) * 2020-11-24 2023-04-28 福建医科大学 PARP protein degradation agent and preparation method and application thereof
US12115154B2 (en) 2020-12-16 2024-10-15 Srx Cardio, Llc Compounds for the modulation of proprotein convertase subtilisin/kexin type 9 (PCSK9)
CN112957463A (en) * 2021-02-22 2021-06-15 深圳市康居正医药科技有限公司 Immune activation type antibody and application thereof
CN117042806A (en) * 2021-03-08 2023-11-10 启德医药科技（苏州）有限公司 Antibody-immune agonist conjugates and their applications
CN115068625B (en) * 2021-03-16 2024-07-02 中国科学院上海药物研究所 PD-L1 and TLR7 double-targeting nano antibody coupled drug and application thereof in resisting tumor
WO2022215704A1 (en) * 2021-04-09 2022-10-13 国立大学法人大阪大学 Ligand-nucleic acid conjugate body
CN114163420B (en) * 2021-10-26 2023-01-24 中山大学附属第一医院 Endoplasmic reticulum Golgi matrix targeting small molecule, conjugate and application thereof
WO2023116413A1 (en) * 2021-12-20 2023-06-29 Versitech Limited Method of developing a peptide-based vaccine conjugated with 1v209
US20250197503A1 (en) * 2022-03-15 2025-06-19 Shanghai Institute Of Materia Medica , Chinese Academy Of Sciences ANTI-HUMAN PD-Ll AND TLR7 DOUBLE-TARGETING NANOBODY CONJUGATE DRUG AND USE THEREOF IN RESISTING TUMOR
CN114984014B (en) * 2022-06-24 2023-09-19 中国人民解放军陆军特色医学中心 Inhibitor for treating sepsis and application thereof
US11779572B1 (en) 2022-09-02 2023-10-10 Deciphera Pharmaceuticals, Llc Methods of treating gastrointestinal stromal tumors
CN116966313A (en) * 2023-04-27 2023-10-31 深圳市康居正医药科技有限公司 Immune activated small molecular compound of coupling antibody and application thereof
WO2025040743A1 (en) * 2023-08-22 2025-02-27 Univerza V Ljubljani Conjugated tlr7 and rig-i agonists

Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102993265A (en) * 2012-10-10 2013-03-27 靳广毅 Immunoreceptor Modulator Conjugate, Its Preparation Method and Application, Its Conjugated Precursor and Synthetic Conjugated Precursor Compound
CN105315281A (en) * 2015-11-16 2016-02-10 深圳大学 Compound I and compound II as well as preparation methods and application thereof

Family Cites Families (11)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN101230064A (en) * 2007-01-22 2008-07-30 靳广毅 9 methylene aryl substituted purine derivative
HUE025555T2 (en) 2007-02-07 2016-02-29 Univ California Conjugates of synthetic tlr agonists and uses therefor
CN101239980B (en) * 2008-02-18 2011-06-22 靳广毅 Immune receptor regulator couplet precursor, couplet and application thereof
EP2396328A2 (en) * 2009-02-11 2011-12-21 The Regents of The University of California Toll-like receptor modulators and treatment of diseases
CN103254304A (en) * 2012-05-30 2013-08-21 深圳大学 Immune stimulant couplet, preparation method thereof and antineoplastic application of the immune stimulant couplet
CN103566377A (en) * 2012-07-18 2014-02-12 上海博笛生物科技有限公司 Targeted immunotherapy for cancer
CN103467590B (en) * 2013-09-02 2016-04-13 深圳大学 Bioconjugation body, the purine compound preparing it, synthetic method, medicinal preparations and its application in immunomodulatory
CN106267188B (en) * 2016-08-15 2025-06-17 深圳大学 A novel antibody combining small molecule immune agonist and PD-1 antibody and its application in anti-tumor
CN107281483B (en) 2017-04-25 2025-10-24 深圳大学 Immune agonist and composition and application thereof and preparation method of the composition
CN107281484A (en) 2017-05-31 2017-10-24 深圳大学 Composition for preparing hepatitis B vaccine and its preparation method and application
CN115192726A (en) * 2018-04-03 2022-10-18 深圳大学 Synthesis of immune agonist targeting compound and application thereof

Patent Citations (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN102993265A (en) * 2012-10-10 2013-03-27 靳广毅 Immunoreceptor Modulator Conjugate, Its Preparation Method and Application, Its Conjugated Precursor and Synthetic Conjugated Precursor Compound
CN105315281A (en) * 2015-11-16 2016-02-10 深圳大学 Compound I and compound II as well as preparation methods and application thereof

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE CAPLUS CHEMICAL ABSTRACTS SERVICE, COLUMBUS, OHIO, US; Registry No. 1429327-81-0, Entered: 26 Apr 2013 (Year: 2013) *
Gao et al., Sci Rep 6, 39598 (2016) (Year: 2016) *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US12172992B2 (en) 2022-03-22 2024-12-24 Abbvie Inc. Pyrimidines for degrading Bruton's tyrosine kinase

Also Published As

Publication number Publication date
CN108379591B (en) 2022-03-29
CN108379591A (en) 2018-08-10
WO2019192454A1 (en) 2019-10-10
US12281117B2 (en) 2025-04-22
US20210038605A1 (en) 2021-02-11
CN115192726A (en) 2022-10-18
JP2021519792A (en) 2021-08-12
WO2019192454A9 (en) 2020-01-16

Similar Documents

Publication Publication Date Title
US12281117B2 (en) Small molecule immune agonists and immune targeting compounds and application thereof
AU2008227128B2 (en) Conjugates of synthetic TLR agonists and uses therefor
US9913834B2 (en) Bis-benzylidine piperidone proteasome inhibitor with anticancer activity
US20220289711A1 (en) Compounds and uses thereof
US20140256922A1 (en) Toll-like receptor-7 and -8 modulatory 1h imidazoquinoline derived compounds
US20180104270A1 (en) Compositions and methods for treating disease states associated with activated t cells and/or b cells
JP2013040209A (en) Tlr agonist
US12485174B2 (en) HDAC6-activated macrophages, compositions, and uses thereof
US20140154202A1 (en) Process for the identification of compounds for treating cancer
AU2013214783A1 (en) CDK8/CDK19 selective inhibitors and their use in anti-metastatic and chemopreventative methods for cancer
US11331304B2 (en) YAP1 inhibitors and methods
CN106659716A (en) Apitimod compositions and methods of use thereof
US20250082763A1 (en) Combination drug for treating triple-negative breast cancer
US20230233696A1 (en) Triantennary n-acetylgalactosamine modified hydroxyl polyamidoamine dendrimers and methods of use thereof
US20230190927A1 (en) Enpp1 inhibitors and methods of modulating immune response
AU2017273871B2 (en) Polymer linkers and their uses
US10266490B2 (en) Radioprotector compounds
US20170035894A1 (en) Drug delivery conjugates for treating resistant cancer and for use in combination therapy
AU2023360058A1 (en) Method of increasing immune cell activation and/or treating cancer using dibenzoxazepinones.
CN113395968A (en) Combination of FAK inhibitors and BTK inhibitors for the treatment of diseases
JP2017501115A (en) DUPA-indenoisoquinoline complex
WO2025188920A1 (en) Local application of agents inhibiting tyrosine kinase or shp-1 signaling
EP4444736A2 (en) Stapled peptide-antibody conjugates (spacs) and uses thereof
KR20230009454A (en) Myristoyl derivatives of 9-amino-doxycycline for cancer stem cell targeting and metastasis prevention
KR20240107169A (en) Nucleic acid compositions and uses thereof

Legal Events

Date Code Title Description
FEPP Fee payment procedure

Free format text: ENTITY STATUS SET TO UNDISCOUNTED (ORIGINAL EVENT CODE: BIG.); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY

STPP Information on status: patent application and granting procedure in general

Free format text: APPLICATION UNDERGOING PREEXAM PROCESSING

AS Assignment

Owner name: SHENZHEN UNIVERSITY, CHINA

Free format text: ASSIGNMENT OF ASSIGNORS INTEREST;ASSIGNORS:JIN, GUANGYI;WANG, ZHULIN;TANG, LI;REEL/FRAME:053958/0321

Effective date: 20200921

FEPP Fee payment procedure

Free format text: ENTITY STATUS SET TO SMALL (ORIGINAL EVENT CODE: SMAL); ENTITY STATUS OF PATENT OWNER: SMALL ENTITY

STPP Information on status: patent application and granting procedure in general

Free format text: APPLICATION DISPATCHED FROM PREEXAM, NOT YET DOCKETED

STPP Information on status: patent application and granting procedure in general

Free format text: DOCKETED NEW CASE - READY FOR EXAMINATION

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NON FINAL ACTION MAILED

STPP Information on status: patent application and granting procedure in general

Free format text: RESPONSE TO NON-FINAL OFFICE ACTION ENTERED AND FORWARDED TO EXAMINER

STPP Information on status: patent application and granting procedure in general

Free format text: NOTICE OF ALLOWANCE MAILED -- APPLICATION RECEIVED IN OFFICE OF PUBLICATIONS

STPP Information on status: patent application and granting procedure in general

Free format text: PUBLICATIONS -- ISSUE FEE PAYMENT VERIFIED

STCF Information on status: patent grant

Free format text: PATENTED CASE