Cancer immunotherapy

Abstract

The present invention relates to cancer immunotherapy. In particular, the present invention provides methods, compounds, compositions and kits for treating and / or preventing cancer. Specifically, provided are methods for the treatment of cancer comprising administering a TLR2 agonist, such as a conjugate of dipalmitoyl-S-glyceryl-cysteine (Pam2Cys) and polyethylene glycol (PEG), more specifically a Pam2Cys-Ser-PEG compound, and an immunostimulatory agent, such as an anti-PD-1, anti-PDL-1, anti-PL-1, or anti-CTLA-4 immunotherapeutic agent.

Description

[0001] This application is a divisional application of Chinese patent application No. 202080061095.1, filed on September 4, 2020, entitled "Cancer Immunotherapy".

[0002] Cross-reference to earlier applications This application claims priority to Australian Provisional Patent Application No. AU 2019903262 (filed on 4 September 2019) and Australian Provisional Patent Application No. AU 2019904864 (filed on 20 December 2019). The entire contents of each of AU 2019903262 and AU 2019904864 are hereby incorporated by reference. Technical Field

[0003] This invention relates to methods, compounds, compositions, and kits for treating and / or preventing cancer. In one aspect, this invention relates to the use of immunotherapy for treating and / or preventing cancer. Background Technology

[0004] Immunotherapy has shown promise in treating cancer because it can slow the growth and spread of cancer cells and help the immune system destroy existing cancer cells. Immunotherapy can help the immune system by stimulating antigen-presenting cells, T cells, or innate cells to trigger and strengthen the immune system, by modulating inhibitory pathways to reduce immunosuppression in the tumor environment, and / or by enhancing adaptive or innate immune responses.

[0005] An example of immunotherapy is checkpoint inhibitors. Currently, checkpoint inhibitors approved by the U.S. Food and Drug Administration (FDA) target cytotoxic T-lymphocyte-associated protein 4 (CTLA-4), programmed cell death receptor 1 (PD-1), or programmed cell death ligand 1 (PD-L1). These checkpoint inhibitors work by preventing cancer cells from escaping the immune system. The first approved drug, ipilimumab, received FDA approval in 2011 for the treatment of metastatic melanoma. Since ipilimumab, five more checkpoint inhibitors have been approved for a total of 14 different indications.

[0006] Between 2015 and 2017, the number of clinical trials using PD-1 and PD-L1 inhibitors increased by nearly 600%. Despite the significant increase in the number of indications for checkpoint inhibitors in recent years, the increase in the beneficial effects of these drugs in terms of patient response percentages has slowed. While clinical responses to PD-1 immunotherapy are positively correlated with tumor PD-L1 expression and other predictive biomarkers such as pre-existing CD8+ T cell infiltration and mutation / neoantigen load, some patients with tumors that do not express PD-L1 have demonstrated a response to PD-1 pathway blockade in other clinical trials. In fact, recent analyses suggest that in the United States, checkpoint inhibitors elicit a response in less than 13% of patients with various types of cancer.

[0007] Therefore, there is still a need for improved immunotherapies to treat cancer.

[0008] Any reference to prior art in this specification is not an admission or implication that such prior art forms part of the general knowledge of any jurisdiction, or that such prior art can reasonably be expected to be understood as being related to and / or combined with other prior art known to those skilled in the art. Summary of the Invention

[0009] The present invention provides a method for treating, preventing or minimizing cancer progression in a subject, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby treating, preventing or minimizing cancer progression in the subject.

[0010] In any aspect of the invention, the TLR2 agonist may be any one as described herein. Preferably, the TLR2 agonist is a compound defined as any one of formulas (I), (IA1), (IA2), (II), (III), (IV), (V), (VI), (VII), (VIII), (IX), (X), (XI), (XII), (XIII), (XIV), (XV), (XVI), (XVII), (XVIII) and (XIX) (collectively referred to herein as formulas (I)-(XIX)).

[0011] In any aspect of the invention, a TLR2 agonist may be a compound comprising a portion A and polyethylene glycol (PEG), wherein portion A is selected from A1' and A2 as defined herein, wherein portion A and PEG are linked by an ester of glycine, serine, homoserine, threonine, phosphoserine, asparagine, or glutamine residues or glutamine residues.

[0012] In any aspect of this invention, the compound may comprise or consist of a portion of the structure A1Y' or A2Y': (A1Y') (A2Y') R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; In each of the cases b, v, w, and z, R 11 R 12 R x R y R 14 R 15 R 16 and R 17 Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19 It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted; and A1Y' or A2Y' is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0013] In some embodiments, the TLR2 agonist is a compound selected from compounds 001-010, A101-A114, and A201-A232.

[0014] As used herein, reference to "compounds of the present invention" may refer to any of the following: • Compounds of formula (I)-(XIX); • A compound selected from any one of compounds 001-010, A101-A114, and A201-A232; • A compound comprising a portion A and polyethylene glycol (PEG), wherein portion A is selected from A1' and A2 as defined herein, wherein portion A and PEG are linked by a glycine, serine, homoserine, threonine, phosphoserine, asparagine, or glutamine residue or an ester of a glutamine residue; and / or • Compounds containing a partial structure of formula (A1Y') or (A2Y') covalently linked to PEG.

[0015] In any aspect of this invention, the immunostimulant may be selected from any of those described herein, including but not limited to the group consisting of: - Cellular immunotherapy (cytotoxic cell immunotherapy or adoptive cell therapy). - Oncolytic virus; - Cancer vaccines; - T-cell conjugate; - Bispecific T-cell conjugates; and - Checkpoint inhibitors.

[0016] In a preferred embodiment, the immunostimulant used according to any of the methods described herein is a checkpoint inhibitor.

[0017] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and a checkpoint inhibitor, thereby treating, preventing, or minimizing cancer progression in the subject.

[0018] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject who has received or is receiving an immunostimulant, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist, thereby treating, preventing, or minimizing cancer progression in the subject.

[0019] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject who has received or is receiving a checkpoint inhibitor, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist, thereby treating, preventing, or minimizing cancer progression in the subject.

[0020] In any aspect of the invention, the TLR2 agonist and the immunostimulant may be administered simultaneously. Alternatively, they may be administered sequentially. For example, the immunostimulant may be administered before the TLR2 agonist, or the TLR2 agonist may be administered before the immunostimulant. Alternatively, treatment with the immunostimulant and / or the TLR2 agonist may be interleaved. In a preferred embodiment, the immunostimulant is a checkpoint inhibitor. In this aspect of the invention, the TLR2 agonist may be administered once or twice a week, and the checkpoint inhibitor may be administered once every three weeks.

[0021] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject, the method comprising the following steps: - Identify subjects with cancer who have received or are currently receiving immunostimulants for cancer treatment. - Assess whether the subject responds to the immunostimulant. -If the subject does not respond to the immunostimulant, administer a therapeutically effective dose of a TLR2 agonist to the subject. This can treat, prevent, or minimize the progression of the subject's cancer.

[0022] In a preferred embodiment, the immunostimulant is a checkpoint inhibitor, preferably a PD-1 inhibitor, a PD-L1 inhibitor, or a CTLA-4 inhibitor. More preferably, the checkpoint inhibitor is a PD-1, PD-L1, or CTLA-4 antibody.

[0023] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject, the method comprising the following steps: - Identify subjects who have cancer and do not respond to treatments containing immunostimulants. -Administer to the subject a therapeutically effective amount of a compound containing, consisting of, or substantially consisting of a TLR2 agonist. This can treat, prevent, or minimize the progression of the subject's cancer.

[0024] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject, the method comprising the following steps: - Identifying subjects with cancer; and - The subject was given a therapeutically effective dose of a TLR2 agonist and an immunostimulant. This can treat, prevent, or minimize the progression of the subject's cancer.

[0025] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject, the method comprising the following steps: - Identifying subjects with cancer; and - The subject was given a therapeutically effective dose of a TLR2 agonist and a checkpoint inhibitor. This can treat, prevent, or minimize the progression of the subject's cancer.

[0026] In any aspect of the invention, TLR2 agonists may be administered in compositional form. Furthermore, in any aspect of the invention, immunostimulants may be administered in compositional form. In a preferred embodiment, checkpoint inhibitors may be administered in compositional form. Typically, the composition further comprises a pharmaceutically acceptable carrier, diluent, or excipient.

[0027] In any embodiment of the invention, the composition may be formulated for intravenous administration to a subject. In other words, the composition is suitable for intravenous administration. In another embodiment of the invention, the composition is formulated for administration over the respiratory tract, preferably by inhalation or intranasal administration. In another preferred embodiment, the composition is formulated as a nasal spray or nasal drops. In another embodiment, a TLR2 agonist is formulated for administration over the respiratory tract, preferably by inhalation, and an immunostimulant, preferably any checkpoint inhibitor described herein, is formulated for intraperitoneal or intravenous administration. In this embodiment, the immunostimulant and checkpoint inhibitor may be administered simultaneously or at different times.

[0028] In one embodiment, the TLR2 agonist is administered in the form of a TLR2 composition, which may not contain any compounds that act as agonists of other TLRs. Preferably, the only TLR agonist present in the TLR2 composition is a TLR2 homodimer or heterodimer agonist. Preferably, the composition contains only one TLR2 agonist.

[0029] In any embodiment, the composition comprises a TLR2 agonist and an immunostimulant, preferably a checkpoint inhibitor, and a pharmaceutically acceptable carrier, diluent, or excipient, substantially comprising or consisting of thereof.

[0030] In another aspect, the present invention further provides a composition comprising a TLR2 agonist and an immunostimulant, preferably a checkpoint inhibitor, and a pharmaceutically acceptable carrier, diluent, or excipient, substantially comprising or consisting of thereof.

[0031] In another aspect, the present invention further provides a method for increasing the survival rate of a subject suffering from cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby increasing the survival rate of the subject suffering from cancer.

[0032] In another aspect, the present invention further provides a method for increasing the survival rate of a subject suffering from cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and a checkpoint inhibitor, thereby increasing the survival rate of the subject suffering from cancer.

[0033] In another aspect, the present invention further provides a method for minimizing, reducing, or preventing tumor growth in a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby minimizing, reducing, or preventing tumor growth in the subject with cancer.

[0034] In another aspect, the present invention further provides a method for minimizing, reducing, or preventing tumor growth in a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and a checkpoint inhibitor, thereby minimizing, reducing, or preventing tumor growth in the subject with cancer.

[0035] In another aspect, the present invention further provides a method for minimizing, reducing, or preventing metastasis in a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby minimizing, reducing, or preventing metastasis in the subject with cancer. In a preferred embodiment, the method minimizes, reduces, or prevents metastasis to the lungs.

[0036] In another aspect, the present invention further provides a method for minimizing, reducing, or preventing metastasis in a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and a checkpoint inhibitor, thereby minimizing, reducing, or preventing metastasis in the subject with cancer. In a preferred embodiment, the method minimizes, reduces, or prevents metastasis to the lungs.

[0037] In any embodiment, the present invention further provides a method for maximally reducing, lowering, or preventing cancer in a subject, the method comprising: - Identifying subjects with tumors capable of metastasis; and - Administer therapeutically effective doses of TLR2 agonists and immunostimulants to the subjects. This would minimize, reduce, or prevent the subject's cancer to the greatest extent possible.

[0038] In any embodiment, the present invention further provides a method for maximally reducing, lowering, or preventing cancer in a subject, the method comprising: - Identifying subjects with tumors capable of metastasis; and -Administer therapeutically effective doses of TLR2 agonists and checkpoint inhibitors to subjects. This would minimize, reduce, or prevent the subject's cancer to the greatest extent possible.

[0039] In any embodiment, the present invention further provides a method for minimizing, reducing, or preventing metastasis in a subject with cancer, the method comprising: - Identify subjects with primary tumors capable of metastasis; -Removal of the primary tumor from the subject; and - Administer therapeutically effective doses of TLR2 agonists and immunostimulants to the subjects. This would minimize, reduce, or prevent metastasis in the subject with cancer.

[0040] In any embodiment, the present invention further provides a method for minimizing, reducing, or preventing metastasis in a subject with cancer, the method comprising: - Identify subjects with primary tumors capable of metastasis; -Removal of the primary tumor from the subject; and -Administer therapeutically effective doses of TLR2 agonists and checkpoint inhibitors to subjects. This would minimize, reduce, or prevent metastasis in the subject with cancer.

[0041] In another aspect, the present invention further provides a method for maximally reducing, lowering or preventing tumor growth in at least one site of a subject, away from the primary tumor site, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby maximally reducing, lowering or preventing tumor growth in at least one site of the subject, away from the primary tumor site.

[0042] In another aspect, the present invention further provides a method for minimizing, reducing, or preventing tumor growth in at least one site of a subject, away from the primary tumor site, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and a checkpoint inhibitor, thereby minimizing, reducing, or preventing tumor growth in at least one site of the subject, away from the primary tumor site.

[0043] In any aspect of the invention, the methods described herein further include identifying a subject suffering from cancer. In one embodiment, the cancer may be precancerous or non-metastatic. In another embodiment, the cancer may be malignant or metastatic.

[0044] In another aspect, the present invention further provides the use of compounds comprising, consisting of, or substantially consisting of, a TLR2 agonist and an immunostimulant in the preparation of a medicament for treating, preventing, or minimizing cancer progression in a subject.

[0045] In another aspect, the present invention further provides the use of compounds comprising, consisting of, or substantially consisting of a TLR2 agonist in the preparation of a first drug and the use of compounds comprising, consisting of, or substantially consisting of an immunostimulant in the preparation of a second drug, wherein the first and second drugs are used for: • To treat, prevent, or minimize cancer progression in the subject. • To minimize, reduce, or prevent tumor growth in the subject. • To minimize, reduce, or prevent subject transfer to the greatest extent possible, or • Increase the survival rate of subjects.

[0046] Alternatively, the first and second drugs may be used in any other method or use of the invention as described herein.

[0047] In another aspect, the present invention further provides the use of TLR2 agonists and immunostimulants for the treatment, prevention, or prevention of cancer progression in subjects.

[0048] In another aspect, the present invention further provides TLR2 agonists and immunostimulants for treating, preventing, or minimizing cancer progression in a subject. Alternatively, TLR2 agonists and checkpoint inhibitors are used in any other method or use of the present invention as described herein, including treating, preventing, or minimizing cancer progression in a subject, minimizing, reducing, or preventing tumor growth in a subject, minimizing, reducing, or preventing metastasis in a subject, or increasing survival in a subject.

[0049] In another aspect, the present invention further provides the use of TLR2 agonists in the preparation of medicaments for: • To treat, prevent, or minimize cancer progression in subjects who have received or are receiving immunostimulants. • To minimize, reduce, or prevent tumor growth in subjects who have received or are receiving immunostimulants. • To minimize, reduce, or prevent metastasis in subjects who have received or are receiving immunostimulants, or • Increase the survival rate of subjects who have received or are receiving immunostimulants.

[0050] In any aspect of the invention, any medicament described herein is suitable for intraperitoneal, intratumoral, topical, oral, or intravenous administration into the respiratory tract, preferably by inhalation or intranasal, subcutaneous, or intramuscular administration. Preferably, any medicament described herein is suitable for intravenous or inhalation administration. In this respect, the medicament may be formulated as a nasal spray or nasal drops.

[0051] In another aspect, the present invention further provides a TLR2 agonist for treating, preventing, or minimizing cancer progression in subjects who have received or are receiving immunostimulants. Alternatively, the TLR2 agonist can be used in any other method or use of the invention as described herein. In one aspect of the invention, the TLR2 agonist used is suitable for intraperitoneal, intratumoral, local, oral, preferably inhalation or intranasal administration to the respiratory tract, intravenous, subcutaneous, or intramuscular routes. Preferably, the TLR2 agonist used is suitable for intravenous or inhalation administration. In another aspect, the TLR2 agonist used can be formulated as a nasal spray or drops for intranasal administration.

[0052] In another aspect, the present invention further provides the use of a TLR2 agonist for the treatment, prevention, or minimization of cancer progression in subjects who have received or are receiving immunostimulants. Alternatively, the use of a TLR2 agonist is in any other method or use of the invention as described herein. In one aspect of the invention, the use of a TLR2 agonist is suitable for intraperitoneal, intratumoral, local, oral, preferably by inhalation or intranasal administration to the respiratory tract, intravenous, subcutaneous, or intramuscular route. Preferably, the use of a TLR2 agonist is suitable for intravenous or inhalation administration. In another aspect, the use of a TLR2 agonist can be formulated as a nasal spray or nasal drops.

[0053] In any embodiment of the present invention, the immunostimulant is selected from the group consisting of: - Cellular immunotherapy (cytotoxic cell immunotherapy or adoptive cell therapy). - Oncolytic virus; - Cancer vaccines; - T-cell conjugate; - Bispecific T-cell conjugates; and - Checkpoint inhibitors.

[0054] In any aspect of this invention, the method does not include the application of: antigen; peptide antigen; or T helper antigen.

[0055] In any aspect of this invention, TLR2 agonists are not administered as part of a vaccine formulation, but are typically administered via subcutaneous, inhalation, intranasal, intradermal, or intramuscular routes.

[0056] In any aspect of this invention, the TLR2 agonist is not administered with the antigen. In another aspect, the TLR2 agonist is not administered with the cell-penetrating peptide.

[0057] In no aspect of this invention is the TLR2 agonist Pam3Cys.

[0058] In any embodiment of the invention, the effects of any TLR2 agonist and immunostimulant described herein may be significant compared to the effects of a single TLR2 agonist or a single immunostimulant. In one aspect, the effects may be additive or synergistic.

[0059] In another aspect, when subjects are administered any of the TLR2 agonists and immunostimulants described herein, the TLR2 agonists may enhance the effectiveness of any of the immunostimulants described herein. Preferably, the enhanced effectiveness of any of the immunostimulants described herein is associated with tumors that are partially or completely resistant to checkpoint inhibitors, such as PD-1 resistant tumors.

[0060] In one embodiment, any TLR2 agonist and immunostimulant described herein may have a significantly greater effect on subject survival than the effect of either TLR2 agonist or immunostimulant alone. In another embodiment, any TLR2 agonist and immunostimulant described herein may have a significantly greater effect on tumor growth or metastasis in subjects than the effect of either TLR2 agonist or immunostimulant alone.

[0061] In any aspect of the invention, the TLR2 agonist and / or immunostimulant is administered once. In another embodiment, the subject is administered the TLR2 agonist and / or immunostimulant two, three, four, or more times.

[0062] In any aspect of the invention, the TLR2 agonist and / or immunostimulant may be administered in the same composition or in separate compositions. In another aspect, the TLR2 agonist and / or immunostimulant may therefore be administered together or sequentially. Alternatively, administration may be staggered. The TLR2 agonist and / or immunostimulant may also be administered at the same frequency or at different frequencies.

[0063] In any aspect of this invention, the TLR2 agonist and immunostimulant can be administered via any route of administration known in the art, including intraperitoneal, intratumoral, local, oral, inhalation or intranasal administration to the respiratory tract, intravenous, subcutaneous or intramuscular routes. Preferably, the TLR2 agonist and / or immunostimulant are administered intravenously or by inhalation.

[0064] In any aspect of the invention, the amount of the applied TLR2 agonist may range from about 250 nmol / kg body weight / dose to 0.005 nmol / kg body weight / dose. Preferably, this range is from about 250 nmol / kg body weight / dose to 0.05 nmol / kg body weight / dose. In some embodiments, the body weight / dose range is from about 250 nmol / kg to 0.1 nmol / kg, from about 50 nmol / kg to 0.1 nmol / kg, from about 5 nmol / kg to 0.1 nmol / kg, from about 2.5 nmol / kg to 0.25 nmol / kg, or from about 0.5 nmol / kg to 0.1 nmol / kg body weight / dose. In some embodiments, the amount is or is about 250 nmol, 50 nmol, 5 nmol, 2.5 nmol, 0.5 nmol, 0.25 nmol, 0.1 nmol, or 0.05 nmol / kg body weight / dose of the compound.

[0065] In any aspect of the invention, the amount of the applied TLR2 agonist may be in the range of about 0.1, 0.2, 0.3, 0.4, 0.5, 0.6, 0.7, 0.8, 0.9, 1.0, 5, 10, 15, 20, 25, 30, 35, 40, 45, 50, 55, 60, 65, 70, 75, 80, 85, 90, 95 or 100 µg / kg or more.

[0066] In any aspect of the invention, the amount of the immunostimulant applied, particularly the amount of the checkpoint inhibitor, can be in the range of about 0.01 mg / kg to about 20 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 5 mg / kg, about 1 mg / kg to about 5 mg / kg, about 2 mg / kg to about 5 mg / kg, about 7.5 mg / kg to about 12.5 mg / kg, or about 0.1 mg / kg to about 30 mg / kg of the subject's body weight. For example, the dose can be about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg, or about 10 mg / kg of body weight, or about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg of body weight.

[0067] In any aspect of the invention, the cancer is selected from the group consisting of: breast cancer, colorectal cancer, adenocarcinoma, mesothelioma, bladder cancer, prostate cancer, germ cell cancer, hepatocellular carcinoma / cholangiocarcinoma, neuroendocrine carcinoma, pituitary tumor, small round cell tumor, squamous cell carcinoma, melanoma, atypical fibroxanthoma, seminoma, non-seminomatous tumor, mesenchymal rheumatoid arthritis, Sertoli cell carcinoma, skin tumor, kidney tumor, testicular tumor, brain tumor, ovarian tumor, gastric tumor, oral cavity tumor, bladder tumor, bone tumor, cervical tumor, esophageal tumor, laryngeal tumor, liver tumor, lung tumor, fibrosarcoma, vaginal tumor, or Wilms' tumor. In a preferred embodiment, the cancer is melanoma, breast cancer, or colon cancer.

[0068] In any aspect of the invention, the checkpoint inhibitor may be a PD-1, PD-L1, or CTLA-4 checkpoint inhibitor. In one aspect, the checkpoint inhibitor is an antibody. Preferably, the checkpoint inhibitor is an antibody form of an inhibitor of PD-1, PD-L1, or CTLA-4.

[0069] Preferably, the TLR2 agonist is any one described herein, and even more preferably Pam2Cys-Ser-PEG. As used herein, Pam2Cys-Ser-PEG can be a compound of the following formula: In another preferred aspect, the TLR2 agonist is a compound of the following formula: In any embodiment of the invention, the method does not involve the administration of an agonist of TLR other than a TLR2 homodimer or heterodimer.

[0070] In one aspect of the invention, administering a therapeutically effective amount of a TLR2 agonist and a therapeutically effective amount of an immunostimulant to a subject comprises administering Pam2Cys-Ser-PEG and the immunostimulant according to the methods described herein. Preferably, the immunostimulant is a checkpoint inhibitor, which is an inhibitor of PD-1, PD-L1, or CTLA-4. More preferably, the checkpoint inhibitor is in the form of an antibody.

[0071] In any aspect of the invention, administering a therapeutically effective amount of a TLR2 agonist and a therapeutically effective amount of a checkpoint inhibitor to a subject comprises administering compound A108 and an immunostimulant according to the methods described herein. Preferably, the immunostimulant is a checkpoint inhibitor that is an inhibitor of PD-1, PD-L1, or CTLA-4. More preferably, the checkpoint inhibitor is in the form of an antibody. In this or any other aspect, the TLR2 agonist and / or checkpoint inhibitor may be administered through the respiratory tract, preferably by inhalation.

[0072] In any aspect of the invention, the TLR2 agonist comprises a lipid, peptidoglycan, lipoprotein, or lipopolysaccharide. Preferably, the TLR2 agonist comprises palmitoyl, myristoyl, stearoyl, lauroyl, capryloyl, or decanoyl. The TLR2 agonist may be selected from the group consisting of Pam2Cys, Pam3Cys, Ste2Cys, Lau2Cys, and Oct2Cys. In a preferred embodiment, the TLR2 agonist comprises Pam2Cys.

[0073] In any aspect of this invention, the TLR2 agonist may be conjugated with other compounds or functional groups. These other compounds or functional groups are any of those described herein. Preferred compounds are selected based on their ability to help dissolve the TLR2 agonist in a carrier, diluent, excipient, or solvent.

[0074] Depending on the polarity of the solvent, the solubility of this TLR2 agonist can be increased by a solubilizer. Therefore, the compound may contain a TLR2 agonist and a solubilizer. Preferably, the TLR2 agonist is linked to the solubilizer. The TLR2 agonist may be polyethylene glycol-modified. Preferably, the solubilizer is any molecule as described herein.

[0075] The solubilizer may contain positively or negatively charged groups, and may consist substantially of or be composed of positively or negatively charged groups. Preferably, the charged groups are branched or linear peptides. Preferably, the positively charged groups contain at least one positively charged amino acid, such as arginine or lysine residues. Preferably, the negatively charged groups contain at least one negatively charged amino acid, such as glutamic acid or aspartic acid. The charged amino acids may be terminal, preferably N-terminated.

[0076] Typically, the solubilizer comprises polyethylene glycol (PEG) or R4. In any aspect of the invention, the solubilizer comprises polyethylene glycol (PEG) and R4.

[0077] In any aspect of the invention, the compound comprises with PEG 11 Conjugated Pam2Cys. In any aspect of this invention, Pam2Cys and PEG 11 The molecules are separated by at least one serine residue.

[0078] In any aspect of the invention, TLR2 agonists exhibit improved solubility compared to other TLR2 agonists. In a preferred embodiment, compound A108 has a solubility approximately 10 times that of PEG-Pam2Cys-R4.

[0079] In one embodiment, compound 1 is present at EC50 concentrations between about 0.2 pg / ml and 500 pg / ml or higher. 50Compound 1 has a stimulatory effect on human TLR2. In a preferred embodiment, compound 1 was used at EC50 concentrations of 0.5 pg / ml, 1.9 pg / ml, 7.8 pg / ml, 31.25 pg / ml, 125 pg / ml and 500 pg / ml. 50 It has a stimulating effect on human TLR2.

[0080] On another note, compound 1 was only found at 10 ng / ml EC 50 The lower TLR2 is stimulated. In other words, compound 1 at 10 ng / ml EC 50 It does not stimulate any other TLR2 agonists.

[0081] In one embodiment, Pam2CysSK4 is present at EC5 concentrations between approximately 0.05 pg / ml and 64 pg / ml or higher. 50 Pam2CysSK4 has a stimulatory effect on human TLR2. In another embodiment, Pam2CysSK4 was found to have a stimulatory effect on human TLR2 at EC2 concentrations of 0.5 pg / ml, 1.9 pg / ml, 7.8 pg / ml, 31.3 pg / ml, 125 pg / ml and 500 pg / ml. 50 Pam2CysSK4 has a stimulatory effect on human TLR2. In another embodiment, Pam2CysSK4 was found to have a stimulatory effect on human TLR2 at EC2 concentrations of 0.0625 pg / ml, 0.25 pg / ml, 1.0 pg / ml, 4 pg / ml, 16 pg / ml and 64 pg / ml. 50 It has a stimulating effect on human TLR2.

[0082] In one embodiment, compound A108 is present at EC50 concentrations between about 0.2 pg / ml and 500 pg / ml or higher. 50 It has a stimulating effect on human TLR2.

[0083] In any aspect of the invention, any TLR2 agonist described herein does not exhibit cytotoxicity. In one aspect, any TLR2 agonist described herein does not inhibit human cytochrome p450 enzymes.

[0084] In another respect, any TLR2 agonist described herein does not significantly increase type I interferon (IFN-α) and type II interferon (IFN-γ). In another respect, any TLR2 agonist described herein is capable of inducing an immune response, preferably by increasing monocyte chemoattractant protein-1 (MCP-1).

[0085] In another aspect, any TLR2 agonist described herein has a half-life between about 1 and 10 hours, preferably between about 3 and 6 hours. In a preferred embodiment, the TLR2 agonist is compound A108.

[0086] In one aspect, any TLR2 agonist described herein can activate the TLR2 pathway by activating either the TLR2 homodimer or the TLR2 / 6 heterodimer. In one aspect, compound 1 has stimulatory effects on both the human TLR2 homodimer and the TLR2 / 6 heterodimer at concentrations above about 1 pg / ml, preferably above about 2 pg / ml. In one aspect, compound A108 has stimulatory effects on the human TLR2 homodimer at concentrations above about 6 pg / ml, preferably above about 7 pg / ml. In one aspect, compound A108 has stimulatory effects on the human TLR2 / 6 heterodimer at concentrations above about 0.3 pg / ml, preferably above about 0.4 pg / ml. In another aspect, any TLR2 agonist described herein does not activate the TLR2 / 1 heterodimer.

[0087] TLR2 agonists considered for use in any aspect of the present invention are any compounds as described herein, including compounds of formula (I)-(XIX).

[0088] In another aspect, the present invention provides a method for treating, preventing, or minimizing cancer progression in a subject, the method comprising administering to the subject a therapeutically effective amount of a compound described herein, such as a compound according to any one of formulas (I)-(XIX) and any other compound containing part A and an immunostimulant, thereby treating, preventing, or minimizing cancer progression in the subject.

[0089] Further aspects of the invention described in the foregoing paragraphs and further embodiments thereof will become clear from the following description, which is given by way of example and with reference to the accompanying drawings. Attached Figure Description

[0090] Figure 1 Compound 1 / Compound A101 enhanced the efficacy of anti-PD1 immunotherapy in the MC38 model. A. Experimental setup: WT mice carrying MC38 were treated intratumorally with a specified dose of Compound 1 along with an anti-PD1 blocking antibody. Tumor growth was measured and monitored over time. B. Kaplan-Meier survival curve mean ± SEM; n = 10 for control, n = 9 for anti-PD1, anti-PD1 + 5 μg Compound 1 and anti-PD1 + 50 μg Compound 1, n = 8 for anti-PD1 + 25 μg Compound 1, statistically significant; log-rank test.

[0091] Figure 2Compound 1 / Compound A101 enhanced the efficacy of anti-PD1 immunotherapy in the B16F10 model. A. Experimental setup: WT mice carrying B16F10 were treated intratumorally with a specified dose of Compound 1 along with an anti-PD1 blocking antibody or a control. Tumor growth was measured and monitored over time. B. Kaplan-Mayer survival curves (n = 10 for control and anti-PD1, and n = 11 for anti-PD1 + 25 μg Compound 1, and n = 12 for anti-PD1 + 50 μg Compound 1; log-rank test).

[0092] Figure 3 Systemic delivery of compound 1 / compound A101 enhanced the efficacy of anti-PD1 immunotherapy. A. Experimental setup: WT mice carrying MC38 were treated intravenously with a specified dose of compound 1 or control and intraperitoneal injection of anti-PD1 or control IgG. B. Tumor growth was measured and monitored over time and evaluated in response to compound 1, anti-PD-1, compound 1, and anti-PD-1 or control IgG.

[0093] Figure 4 Combination immunotherapy reduced the metastatic spread of 4T1.2 breast cancer cells. A. Experimental setup: Balb / c mice were injected with 4T1.2 breast cancer cells. Primary tumors were surgically removed and the mice were treated as shown in the figure. The number of metastases on the lung surface was counted. B. Number of 4T1.2 lung metastases in the specified cohort (mean ± SEM; n = 8 for control and anti-PD1; n = 9 for anti-PD1 + 10 μg compound 1; statistics, one-way ANOVA with multiple comparisons performed using Dunnettes' test).

[0094] Figure 5 The combination of compound A108 and a checkpoint inhibitor significantly reduced the growth rate of EMT6.5 tumors in vivo. A. Longitudinal monitoring of EMT6.5 tumors (caliper measurements) showed that treatment regimens containing compound A108 reduced EMT6.5 tumor growth and size. Mediator versus anti-PD1 monotherapy (200 μg), p = ns; mediator versus INNA-042 (10 μg), p = 0.0001; mediator versus INNA-042 + anti-PD1, p < 0.0001; anti-PD1 monotherapy (200 μg) versus INNA-042 + anti-PD1, p < 0.0001. B. Kaplan-Meyer survival plots showed better prognosis in animals treated with the combination of compound A108 and anti-PD1. Log-rank test. C. Violin plots detail the size of individual tumors (black dots) in each group.

[0095] Figure 6The combination of compound A108 and a checkpoint inhibitor exhibited antitumor activity in an MC38 model rich in immune cells. WT mice carrying MC38 melanoma were treated intratumorally with 25 μg of compound A108. Kaplan-Mayer survival analysis highlighted a significant improvement in survival in response to administration of compound A108 and anti-PD1 immunotherapy.

[0096] Figure 7 The combination of compound A108 and a checkpoint inhibitor exhibited antitumor activity against large tumors in the MC38 colon cancer model. Compound A108 was administered intratumorally every two days, followed by intraperitoneal administration of anti-PD-1 (10 mg / kg, 200 μg dose) every four days. Tumor growth was measured and monitored over time. Data show the antitumor effects of compound A108, anti-PD-1, or the combination during treatment. Two-way ANOVA with Tukey's multiple comparison test was used.

[0097] Figure 8 Compound A108, in combination with anti-PD-1, anti-PDL1, or anti-CTL4, prevented tumor growth in the WEHI164 fibroblastoma tumor model. A single 2.5 μg dose of compound A108, administered intraperitoneally on day 7 post-inoculation, was combined with A. three doses of anti-PD-1 (200 μg intraperitoneally), B. anti-CTLA4 (100 μg intraperitoneally on day 7), or C. anti-PDL1. A. Anti-PD1 and anti-PD1 + A108 p = 0.034; anti-PD1 + A108 and PBS p = 0.0081; B. Anti-CTLA4 + A108 and anti-CTLA4 p = 0.0237; anti-CTLA4 + A108 and A108 alone p = 0.0001; anti-CTLA4 alone and PBS p = 0.0453; anti-CTLA4 + A108 and PBS p = 0.0001; C. Anti-PDL1 + A108 and A108 alone p = 0.0001; anti-PDL1 alone and PBS p = 0.0414; anti-PDL1 + A108 and PBS p = 0.0028.

[0098] Figure 9 Human TLR2 activity. Human TLR2 activity of compounds A107, A108, A115, A116, A203, A204, A215 and A216 as determined by NK-κB luciferase as described in Example 5. Detailed Implementation

[0099] Referring now to certain embodiments of the invention. While the invention will be described in conjunction with embodiments, it should be understood that it is not intended to limit the invention to those embodiments. Rather, the invention is intended to cover all alternatives, modifications, and equivalents that may be included within the scope of the invention as defined in the claims.

[0100] Those skilled in the art will recognize that many methods and materials similar to or equivalent to those described herein can be used in the practice of this invention. The invention is by no means limited to the methods and materials described herein. It should be understood that the invention disclosed and defined herein is extendable to all alternative combinations of two or more of the individual features mentioned or readily apparent from the textual and accompanying drawings. All these different combinations constitute multiple alternative aspects of the invention.

[0101] All patents and publications mentioned in this article are incorporated herein by reference in their entirety.

[0102] For the purposes of interpreting this specification, terms used in the singular also include the plural and vice versa.

[0103] During tumor development, tumor cells are more or less resistant to the patient's own immune system because they are the patient's own cells (e.g., they are their own) and cannot be effectively recognized by the patient's immune system, allowing tumor cells to grow and divide without proper regulation and control. Therefore, the patient's own immune system needs stimulation to attack cancer cells. Cancer immunotherapy involves utilizing the cancer patient's immune system by stimulating it to reject cancer. In turn, the activated immune system attacks cancer cells without attacking the patient's normal cells. So-called immunotherapies that have proven effective in treating cancer use checkpoint inhibitors.

[0104] Although checkpoint inhibitor cancer immunotherapy has shown efficacy in some cancers, it is ineffective in a significant proportion of patients, and some initial responders eventually develop resistance and relapse. A patient's responsiveness to immunotherapy depends on many factors, including individual genetic makeup, history of infection, age, nutritional status, HLA type, and the consumption of certain medications. Masking tumor antigens, making tumor cells undetectable by immune surveillance, is also a particular problem, as the immune status of cancer patients is often compromised.

[0105] For example, PD-1 blockade alone has been shown to be ineffective in certain patient subgroups with certain types of cancer, such as melanoma and large B-cell lymphoma. Therefore, there is a need for more reliable and effective immunotherapy regimens for treating cancer. The inventors unexpectedly discovered a positive response to immunotherapy when an immunostimulant (e.g., a checkpoint inhibitor) is administered in combination with a TLR2 agonist. This effect has been tested in numerous cancer models, including colon cancer, breast cancer, metastatic breast cancer, fibrosarcoma, and melanoma. Importantly, in some respects, administration of an immunostimulant (e.g., a checkpoint inhibitor) and a TLR2 agonist significantly improves the cancer in subjects.

[0106] Specifically, when subjects were administered a combination of an immunostimulant and a TLR2 agonist, this resulted in a significant inhibition of tumor growth. This effect was observed even at low doses of a given TLR2 agonist. Importantly, when subjects were administered a combination of an immunostimulant and a TLR2 agonist, this resulted in increased survival compared to the effect of administering an immunostimulant alone. In some cases, the inventors found that while administering an immunostimulant alone did not increase subject survival, survival was significantly increased when the immunostimulant was administered in the presence of a TLR2 agonist. Specifically, in melanoma and breast cancer models, a positive response was observed when a checkpoint inhibitor was administered in combination with a TLR2 agonist, although... alone The effect of checkpoint inhibitors is poor or ineffective.

[0107] The inventors have demonstrated this significant effect in many cancer models with different etiologies and pathogenesis.

[0108] Specifically, the combination of TLR agonists and immunostimulants, as described in this article, has been tested in numerous cancer models, including: • Epithelial breast cancer with characteristics of triple-negative breast cancer; • Metastatic PD1-insensitive breast cancer; • Colon cancer; • PD1-insensitive melanoma; and • Fibrosarcoma.

[0109] Therefore, those skilled in the art will understand the applicability of this invention to any other cancers described herein. The findings described herein are significant because they establish that tumors previously untreatable using checkpoint inhibitors alone are now treatable when using a combination of immunostimulants and TLR2 agonists.

[0110] The inventors also describe herein the efficacy of various combinations of TLR2 agonists with various immunostimulants for the treatment of cancer, including: • Compound 1 / compound A101, in models of colon cancer, melanoma, and metastatic breast cancer, when administered intraperitoneally, intratumorally, or systemically, can impair tumor growth, prolong survival, and reduce metastasis; and • Compound A108, in models of colon cancer, breast cancer, melanoma, fibrosarcoma, and metastatic breast cancer, significantly reduced tumor growth, improved survival, and decreased metastasis when administered intraperitoneally, intratumorally, or systemically.

[0111] Therefore, those skilled in the art will understand the applicability of this invention to any other TLR2 agonists described herein.

[0112] The inventors have also demonstrated the efficacy of combining numerous checkpoint inhibitors with the aforementioned TLR2 agonists in the treatment of cancer. These checkpoint inhibitors include: •PD1; •PD-L1; and •CTLA-4.

[0113] Therefore, those skilled in the art will understand the applicability of this invention to any other immunostimulants described herein or known in the art.

[0114] Therefore, this work identifies a novel immunotherapy that can improve response rates in cancer patients, including those who previously did not respond to immune stimulation.

[0115] This effect is surprising to the inventors, as the role of TLR2 agonists in cancer treatment has been unclear. Specifically, subcutaneous or intraperitoneal administration of synthetic TLR2 / 6 agonists (including Pam2cysSK4 and MALP2) has been reported to have no antitumor activity and instead induce IL-10 and Treg (Yamazaki et al. PLOS ONE [PLOS ONE] 2011 6 (4): e18833). The same group reported that intravenous administration of Pam2cysSK4 (a TLR2 / 6 agonist) promoted myeloid-derived suppressor cells (Maruyama et al. Biochemical and Biophysical Research Communications [Biochemical and Biophysical Research Communications] 2015 457: 445e450). Importantly, it has been proposed that endogenous TLR2 / 6 agonists derived from cancer cells may enhance metastasis (Kim et al. Nature [Nature] 2009 457: 102-106). Another study also showed that TLR2 stimulation may promote the growth of colorectal cancer cells through the PI3K / Akt and NFκB signaling pathways (Liu et al., International Immunopharmacology, 2018, 59: 375-383).

[0116] Toll-like receptors (TLRs) Toll-like receptors (TLRs) are pattern recognition receptors (PRRs) expressed by different cell types that play important roles in both innate and adaptive immunity. Cells of the innate immune system respond to TLR activation by producing pro-inflammatory cytokines and chemokines, which send signals to clear pathogens and damage themselves. Upon binding to specific ligands, TLR activation regulates cytokine expression by inducing several adaptor molecules of interferon-β TRIF, including the myeloid differentiation primary response gene 88 (MyD88), the adaptor protein TIRAP containing a Toll-interleukin 1 receptor (TIR) ​​domain, and other adaptors containing TIR domains. This induces activation of transcription factors such as nuclear factor κB (NF)-κB, activator protein-1 (AP-1), and interferon regulatory factor (IRF).

[0117] There are many TLRs belonging to this membrane receptor protein family, including TLR1, TLR3, TLR4, TLR5, TLR6, TLR7, TLR8 and TLR9.

[0118] As used herein, the term "TLR2" is intended to refer to the Toll-like receptor 2 protein. In humans, TLR2 is composed of... TLR2Gene-encoded. TLR2 is expressed on the surface of certain cells and plays a fundamental role in pathogen recognition and innate immune activation.

[0119] TLR2 agonists are agents that bind to Toll-like receptor 2. TLR2 agonists can bind to and activate TLR2 as homodimers or heterodimers. Any TLR2 agonists known in the art are considered for use in this invention.

[0120] In any embodiment of the invention, the TLR2 agonist comprises a lipid, peptidoglycan, lipoprotein, or lipopolysaccharide. Preferably, the TLR2 agonist comprises palmitoyl, myristoyl, stearoyl, lauroyl, capryloyl, or decanoyl. The TLR2 agonist may be selected from the group consisting of Pam2Cys, Pam3Cys, Ste2Cys, Lau2Cys, and Oct2Cys. In a preferred embodiment, the TLR2 agonist comprises Pam2Cys.

[0121] An exemplary lipopeptide according to any embodiment of the present invention is the lipopeptide "Pam2Cys". Those skilled in the art will understand that the term "lipopeptide" refers to any composition of substances comprising one or more conjugated lipid moieties and one or more amino acid sequences. Pam2Cys (also known as dipalmitoyl-S-glycero-cysteine ​​or S-[2,3-bis(palmitoyloxy)propyl]cysteine) has been synthesized, and it corresponds to the lipid moieties of MALP-2, which is derived from fermentation mycoplasma (… Mycoplasma ƒermentans Pam2Cys is a macrophage-activated lipopeptide isolated from TLR2. Pam2Cys is considered a ligand for TLR2.

[0122] Pam2Cys has the following structure: As used in this article, the reference to "S" in the above chemical structure defines a sulfur atom.

[0123] Another exemplary lipopeptide is the lipoamino acid N-palmitoyl-S-[2,3-bis(palmitoyloxy)propyl]cysteine, also known as Pam3Cys, or Pam3Cys-OH, which is the synthetic form of the N-terminal portion of Braun's lipoprotein that spans the inner and outer membranes of Gram-negative bacteria. Pam3Cys has the following structure: US Patent No. 5,700,910 describes several N-acyl-S-(2-hydroxyalkyl)cysteines used as intermediates in the preparation of lipopeptides for use as synthetic adjuvants, B lymphocyte stimulants, macrophage stimulants, or synthetic vaccines. US 5,700,910 also imparts the use of such compounds as intermediates in the synthesis of Pam3Cys-OH and lipopeptides containing this lipoamino acid or its analogue at the N-terminus.

[0124] Other lipid moieties that can be used to target cell surface TLRs include palmitoyl, myristoyl, stearoyl, lauroyl, capryloyl, or decanoyl.

[0125] In addition to Pam2Cys and Pam3Cys, the present invention also contemplates the use of Ste2Cys, Lau2Cys, and Oct2Cys according to the present invention. Those skilled in the art will recognize that Ste2Cys is also called S-[2,3-bis(stearoyloxy)propyl]cysteine ​​or distearyl-S-glycero-cysteine; Lau2Cys is also called S-[2,3-bis(lauroyloxy)propyl]cysteine ​​or dilauroyl-S-glycero-cysteine; and Oct2Cys is also called S-[2,3-bis(octanoyloxy)propyl]cysteine ​​or dioctanoyl-S-glycero-cysteine.

[0126] Other suitable TLR2 agonists include, but are not limited to, synthetic triacylated and diacylated lipopeptides, FSL-1 (derived from Mycoplasma salicifolia), and others. Mycoplasma salivarium ) 1 The synthetic lipoproteins), Pam3Cys (tripalpalmitoyl-S-glycerocysteine) and S-[2,3-bis(palmitoyloxy)-(2RS)-propyl]-N-palmitoyl-(R)-cysteine, where "Pam3" stands for "tripalpalmitoyl-S-glycero". Derivatives of Pam3Cys are also suitable TLR2 agonists, including but not limited to: S-[2,3-bis(palmitoyloxy)-(2-R,S)-propyl]-N-palmitoyl-(R)-Cys-(S)-Ser-(Lys)4-hydroxytrihydrochloride, Pam3Cys-Ser-Ser-Asn-Ala, Pam3Cys-Ser-(Lys)4, Pam3Cys-Ala-Gly, Pam3Cys-Ser-Gly, Pam3Cys-Ser, Pam3Cys-OMe, Pam3Cys-OH, PamCAG, palmitoyl-Cys((RS)-2,3-bis(palmitoyloxy)-propyl)-Ala-Gly-OH, etc.

[0127] Other non-limiting examples of suitable TLR2 agonists are Pam2CSK4, Pam2CysSK4 (dispalmitoyl-S-glycerocysteine-serine-(lysine)4, or Pam2Cys-Ser-(Lys)4), which are synthetic diacylated lipopeptides. Other synthetic TLR agonists include those described, for example, in the following literature: Kellner et al. (1992) Biol. Chem. 373: 1: 51-5; Seifer et al. (1990) Biochem. J. 26: 795-802; and Lee et al. (2003) J. Lipid Res. 44: 479-486.

[0128] TLR2 agonists can be conjugated with one or more compounds or functional groups. Examples of specific compounds or functional groups are given below. One form of compound or functional group can be used to increase the solubility of the TLR2 agonist. As will be understood by those skilled in the art, TLR2 agonists are typically nonpolar, and therefore, while soluble in nonpolar solvents, they dissolve only in small amounts in polar and aqueous solvents. When it is desired to use the TLR2 agonist in polar or aqueous solvents, the TLR2 agonist can be conjugated with a solubilizer.

[0129] The solubilizer may include one or more solubilizers that can be conjugated with a TLR2 agonist to improve the solubility of the TLR2 moiety. The solubilizer is typically a polar portion that increases the solubility of the TLR2 moiety in polar or aqueous solvents.

[0130] In any aspect of this invention, the solubilizer may be a positively charged group. Positively charged groups of this invention include, but are not limited to, transmembrane peptides, HIV Tat 48-60, HIV Rev 34-50, transportans, oligo-arginine peptides (straight-chain and branched-chain), oligo-lysine peptides, pyrrrochoricin, α-helical amphiphilic model peptides, polylysine, protamine, FL17, Magnafloc 1697, and polycationic compounds described in US 6,689,478 and US 4,035,558.

[0131] In another embodiment of the invention, the solubilizer comprises, or is substantially composed of, or is composed of, straight-chain or branched-chain peptides. Typically, the straight-chain or branched-chain peptides comprise positively or negatively charged amino acids. Positively charged amino acids may be lysine, arginine, histidine, ornithine, or combinations thereof. Branched-chain or straight-chain peptides may contain at least one lysine or arginine residue. Preferably, the charged amino acid is terminal, for example, N-terminus. Branched-chain peptides may have one of the following structures. In the above structure, X can be an independently charged residue (positively or negatively charged). Preferably, the positively charged amino acid is lysine, arginine, histidine, or ornithine. Preferably, the negatively charged amino acid is glutamic acid or aspartic acid.

[0132] As used herein, “PEG” refers to the polymer compound polyethylene glycol. Unless otherwise defined, references to “PEG” include ethylene oxide polymers of any length. References to PEG also include substituted PEGs.

[0133] Compounds or functional groups that can be used as solubilizers can be one or more of the group consisting of "PEG" (or polyethylene glycol) and a polar polypeptide, such as "R4", a hyperbranched tetraarginine complex; "H4", a hyperbranched tetrahistine complex; "H8", a linear peptide containing a histidine residue; and "E8", a linear peptide containing a glutamate residue. Other linear and branched lipid solubilizers are also contemplated, including hyperbranched peptides containing glutamate residues (see, for example, "branched E8" below). In yet another embodiment of the invention, the solubilizer comprises PEG and one or more of the group consisting of R4, H4, H8, and E8 (linear or branched). R4, H4, H8, and E8 have previously been described in PCT / AU 2009 / 000469 (WO / 2010 / 115230) and have the following structures: Example R4 Example H4 Example H8 Example E8 Example branch E8 Exemplary PEG The following are schematic diagrams of some examples of branched (structures 1-5) and straight (structures 6-8) immunogenic compositions, which contain positively charged (arginine, R; lysine, K) or negatively charged (aspartic acid, D; glutamic acid, E) amino acids at their terminal positions, such that their respective electrostatic charges are displayed in the environment. Each immunogenic composition also contains a dipalmitoyl group. S- Glycerylcysteine ​​(Pam2Cys) is the ligand for Toll-like receptor 2. Two serine residues (Ser) are also incorporated. For construct 2, the peptide structure is assembled in the N→C direction, while all other structures shown in the figure are assembled in the C→N direction. Positive and negative electrostatic charges are indicated as 2-, 2+, 1-, 1+, etc., according to the magnitude of the charge. Ac = The acetyl group, for example, is used to suppress the positive charge of the α-amino group in relation to the N-terminal glutamate. Those skilled in the art will understand that the present invention is not limited to specific exemplary compounds or functional groups that can be used as solubilizers, and other suitable compounds or functional groups (including those that can be used as solubilizers known in the art) can also be used according to the present invention, such as carbohydrates.

[0134] One or more compounds or functional groups (such as solubilizers) can be conjugated with lipids according to the present invention in a manner well known to those skilled in the art. For example, conjugation is contemplated via Fmoc chemistry, through disulfide or thioether bridges, or via oxime chemistry. In a particular embodiment of the invention, a soluble form of Pam2Cys is prepared by adding O-(N-Fmoc-2-aminoethyl)-O'-(2-carboxyethyl)-undecanediol (Fmoc-PEOn-OH, Merck Ltd) to Pam2Cys. This results in the formation of a PEGylated lipid form, Pam2Cys-PEG. 11 It is then suitable for administration to the subject.

[0135] In another form of the invention, the TLR2 portion comprises a conjugate containing Pam2Cys conjugated with the dangling R4 form. In a preferred form, the dangling-Pam2Cys is conjugated with R4 according to the following structure: In a preferred form of any embodiment of the invention, the TLR2 portion comprises a conjugate containing Pam2Cys conjugated with PEG. 11 or PEG 12 Conjugated Pam2Cys. Preferably, Pam2Cys is conjugated with PEG. 11 or PEG 12 The molecule is separated by at least two serine residues (PEG). 11 -SS-Pam2Cys or PEG 12 -SS-Pam2Cys).

[0136] As used in this article, references to TLR2 agonists also include their pharmaceutically acceptable salts, solvates, polymorphs, or prodrugs.

[0137] Other compounds containing TLR2 agonists that can be used in any aspect of the present invention are described below.

[0138] In any respect, the compound can be a compound of formula (I): Where A contains or is composed of parts selected from A1 and A2: in Each z is independently selected from 1 or 2; Each X is independently selected from -S-, -S(=O)-, and -S(=O)2-; In part of A1: Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R9 and R 10 Independently selected from the following groups: -NH-, -O-, or single bonds; and In part of A2: b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, S(=O), -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O-, -C(=O)S-, -SC(=O-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR- and -NRC(=O)NR-; R 11 R 12 R x R y R 14 R 15 R 16 and R 17 Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19 It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C 20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any one of L1, L2 and L3 is optionally substituted; Y is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; and B contains or is composed of polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0139] In any respect, the compound can be a compound of formula (IA1): Where A contains a portion of A1 or is composed of it: A1 Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; Y is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; and B contains or is composed of polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0140] In some embodiments, g is an integer from 12 to 16.

[0141] In some embodiments, g is 14.

[0142] In any respect, the compound can be a compound of formula (IA2): Where A contains or is composed of the following structures: A2 in b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, S(=O), -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O-, -C(=O)S-, -SC(=O-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR- and -NRC(=O)NR-; R 11R 12 R x R y R 14 R 15 R 16 and R 17 Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19 It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C 20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any one of L1, L2 and L3 is optionally substituted; Y is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; and B contains or is composed of polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0143] In some embodiments, v is an integer selected from 2, 3, 4, or 5. In some embodiments, v is 2 or 3. In some embodiments, v is 2.

[0144] In some embodiments, R x R y R 11 R 12 R 13 R 14 R 15 R 16 and R 17 It is H.

[0145] In some embodiments, R and R 13 Each is H.

[0146] In some embodiments, Z1 and Z2 are the same and are selected from the group consisting of: -O-, -NR-, -S-, S(=O), S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, OC(=O)O-, NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-.

[0147] In some embodiments, Z1 and Z2 are independently selected from the group consisting of: -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-.

[0148] In some embodiments, w is an integer selected from 1 to 7. In some embodiments, w is 1.

[0149] In some embodiments, b is 0.

[0150] In some embodiments, the sum of b and w is 1 to 7. In these embodiments, b can be an integer selected from 0 to 7, and w can be an integer selected from 1 to 7, preferably 1.

[0151] In some embodiments, b is 0, w is 1, and v is 2.

[0152] In some embodiments, R 18 It is H.

[0153] In some embodiments, R 19 Choose from the following groups: H, C1-C6 alkyl, -C(=O)C1-C6 alkyl, or -C(=O)C 11 -C 19 alkyl.

[0154] In some embodiments, R19 It is selected from H, C1-C6 alkyl, -C(=O)C1-C6 alkyl, preferably H, C1-C4 alkyl, -C(=O)C1-C4 alkyl.

[0155] In some embodiments, R 19 Selected from H and -C(=O)CH3.

[0156] In some embodiments, L1 and L2 are independently selected from C5-C 21 Aliphatic groups or C4-C 20 Heteroaliphatic groups. In some embodiments, L1 and L2 are independently selected from C 10 -C 18 Aliphatic group or C 10 -C 18 Heteroaliphatic groups. In some embodiments, L1 and L2 are independently selected from C 14 -alkyl and C 15 -alkyl.

[0157] In some embodiments, X is S.

[0158] In some embodiments, X is S (=O).

[0159] In some embodiments, X is S(=O)2.

[0160] In some embodiments, R6 and R7 are each H.

[0161] In some embodiments, R 18 and R 19 Each is H.

[0162] In some embodiments, the present invention provides a compound of formula (I), wherein: v is an integer from 2 to 5; b is 0; R x R y R 13 R 14 R 15 R 16 and R 17 It is H; Z1 and Z2 are independently selected from the following groups: -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; w is an integer from 1 to 7; R 19Choose from the following groups: H, C1-C6 alkyl, -C(=O)C1-C6 alkyl, or -C(=O)C 11 -C 19 Alkyl; and L1 and L2 are independently selected from C 10 -C 18 Aliphatic group or C 10 -C 18 Aliphatic groups.

[0163] In some embodiments, the present invention provides a compound, wherein v is 2; b is 0; w is 1; The sum of v, b, and w is 3; The sum of b and w is 1; z is 1; X is S Z1 and Z2 are independently selected from the following groups: -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR- and -NRC(=O)NR-; R 11 R 12 R x R y R 14 R 15 R 16 and R 17 In each of the cases b, v, w, and z, H is present. R and R 13 Each is H; R 18 It is H; R 19 Choose from the following groups: H, C1-C6 alkyl, -C(=O)C1-C6 alkyl, or -C(=O)C 11 -C 19 Alkyl; and L1 and L2 are independently selected from C 10 -C 18 Aliphatic group or C 10 -C 18 Aliphatic groups.

[0164] It should be understood that any embodiment of the substituents described herein includes substituents R1, R2, R4, R5, R6, R7, R9, R 10 z, X, g, R 11 R12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y L1, L2, Z1, Z2, b, v, w, n, m, p, q, R3, L, t, k, and h are intended to be applied to any case of substituents in any compound described herein, including compounds of formula (I)-(XIX).

[0165] In any respect, the compound can be a compound of formula (II): Where A includes or consists of A1 or A2 as defined herein; Y' is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; and B contains or is composed of polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0166] In some embodiments, the compound comprises a portion A1, wherein: Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; z is 1; X is S; R6 and R7 are H; R9 and R 10 All are single keys.

[0167] In some embodiments, part A1 is defined by part A1'. A1' Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18.

[0168] In any respect, any compound described herein may be a compound comprising a portion A and PEG, wherein portion A is selected from A1' and A2 as defined herein, wherein portion A and PEG are linked by an ester of glycine, serine, homoserine, threonine, phosphoserine, asparagine, or glutamine residues or glutamine residues.

[0169] In any respect, the compound may contain or consist of partial structures A1Y' or A2Y': (A1Y') (A2Y') R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; In each of the cases b, v, w, and z, R 11 R 12 R x R y R 14 R 15 R 16 and R 17Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19 It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C 20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any of L1, L2, and L3 is optionally substituted; and A1Y' or A2Y' is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0170] In some embodiments, portions A and PEG are linked by serine, homoserine, threonine, or phosphoserine residues.

[0171] In some embodiments, portions A and PEG are obtained by... The bond represents a covalently linked ester of glycine, serine, homoserine, threonine, phosphoserine, asparagine, or glutamine residues.

[0172] In any respect, a compound can be: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; and X is selected from -S-, -S(=O)- and -S(=O)2-; The above structure is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0173] In some embodiments, the compound may be: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; and X is selected from -S-, -S(=O)- and -S(=O)2-; The above structure is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0174] In some embodiments, the compound may be: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are H; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 All are single keys; z is 1; and X is S; The above structure is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0175] In some embodiments, PEG is transmitted by... The bond represents a covalent connection.

[0176] In some embodiments, the compound may be: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; R6 and R7 are H; R9 and R 10 All are single keys; z is 1; and X is S; The above structure is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0177] In some embodiments, PEG is transmitted by... The bond represents a covalent connection.

[0178] In some embodiments, the compound may be: Its salt, solvate, or prodrug, wherein R1, R2, and g are as defined herein.

[0179] In some embodiments, PEG is transmitted by... The bond represents a covalent connection.

[0180] In some embodiments, the compound may be: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; In each of the cases b, v, w, and z, R 11 R 12 R x R y R 14 R 15 R 16 and R 17 Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19 It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C 20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any one of L1, L2 and L3 is optionally substituted; The above structure is covalently linked to polyethylene glycol (PEG). Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0181] In some embodiments, PEG is transmitted by... The bond represents a covalent connection.

[0182] In any respect, the compound can be a compound of formula (III): in AY includes or consists of portions selected from AY1 and AY2. Among them, R1, R2, R6, R7, R9, R 10 z, X, g, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Each of L1, L2, Z1, Z2, b, v, and w is as defined for compounds of formula (I); and B contains or is composed of polyethylene glycol (PEG).

[0183] In any respect, the compound can be a compound of formula (IV): (IV) in n is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0184] In any respect, the compound can be a compound of formula (V): (V) in n is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; q is zero or 1; R1 and R2 are independently chosen from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0185] In some embodiments, the compound is a compound of formula (IV) or (V), wherein R6 and R7 are H; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 All are single keys; z is 1; and X is S.

[0186] In some embodiments, any compound of formula (I)-(V) can be a compound of formula (VI): (VI) in n is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; q is zero or 1; R1 and R2 are independently chosen from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0187] In any respect, the compound can be a compound of formula (VII): (VII) in n is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid; b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; In each of the cases b, v, w, and z, R 11 R 12 R x R y R 14 R 15 R 16 and R 17 Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C 20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any one of L1, L2 and L3 is optionally substituted; Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0188] In any respect, the compound can be a compound of formula (VIII): in A is selected from A1 and A2 as defined in this article. Y is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; n is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; q is zero or 1; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0189] In any respect, the compound can be a compound of formula (IX): in A1 is represented by the part A1 defined by equation (I). Y is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; n is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; q is zero or 1; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0190] In some embodiments, the compound is a compound of formula (VIII) or (IX), wherein R6 and R7 are H; R9 and R 10 All are single keys; z is 1; X is S.

[0191] In any respect, the compound can be a compound of formula (X): in Pam2Cys has the following structure: ; Y is: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; n is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; When q = 1, R3 is H, -NH2, or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0192] In any respect, the compound can be a compound of formula (XI): in Pam2Cys has the following structure: ; Y is: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; n is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; When q = 1, R3 is H, -NH2, or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0193] In any respect, the compound can be a compound of formula (XII): in Pam2Cys has the following structure: ; Y is: R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; n is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; When q = 1, R3 is H, -NH2, or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0194] In any respect, the compound can be a compound of formula (XIII): in Pam2Cys-Ser has the following structure: ; n is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0195] In one embodiment, the compound has the formula (XIV): (XIV) in n is between 3 and 100; k is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; t is 2, 3, or 4; h is 1, 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0196] In one embodiment, the compound has the formula (XV): (XV) in n is between 3 and 100; k is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; t is 2, 3, or 4; h is 1, 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid; b and w are each independent integers from 0 to 7, and v is an integer from 0 to 5, provided that: The sum of b, v, and w is at least 3; and The sum of b and w ranges from 0 to 7; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; Z1 and Z2 are each independently selected from the following groups: -O-, -NR-, -S-, -S(=O)-, -S(=O)2-, -C(=O)O-, -OC(=O)-, -C(=O)NR-, -NRC(=O)-, -C(=O)S-, -SC(=O)-, -OC(=O)O-, -NRC(=O)O-, -OC(=O)NR-, and -NRC(=O)NR-; In each of the cases b, v, w, and z, R 11 R 12 R x R y R 14 R 15 R 16 and R 17 Each is independently an H or C1-C6 aliphatic group; R, R 13 and R 18 Each is independently an H or C1-C6 aliphatic group; R 19 It is H, C1-C6 aliphatic group, amino protecting group, L3-C(=O)-, or A2; L1 and L2 are each independently C5-C 21 Aliphatic groups or C4-C 20 Aliphatic groups; L3 is C1-C 21 Aliphatic group or C2-C 20 Aliphatic groups; A2 is an amino acid or peptide; Among them exist in R, R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 R x R y Any aliphatic or heteroaliphatic group in any one of L1, L2 and L3 is optionally substituted; Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0197] In any respect, the compound can be a compound of formula (XVI): (XVI) in n is between 3 and 100; k is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; t is 2, 3, or 4; h is 1, 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; R6 and R7 are independently selected from the group consisting of: H, straight-chain or branched C1-C4 alkyl groups and -C(=O)CH3; R9 and R 10 It can be independently selected from the following groups: -NH-, -O-, or single bonds; z is 1 or 2; X is selected from -S-, -S(=O)- and -S(=O)2-; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0198] In any respect, the compound can be a compound of formula (XVII): (XVII) in n is between 3 and 100; k is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; t is 2, 3, or 4; h is 1, 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R6 and R7 are H; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; R9 and R 10 All are single keys; z is 1; X is S; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0199] In any respect, the compound can be a compound of formula (XVIII): (XVIII) in n is between 3 and 100; k is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; t is 2, 3, or 4; h is 1, 2, 3, or 4; q is zero or 1; R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; R6 and R7 are H; R9 and R 10 All are single keys; z is 1; X is S; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0200] In any respect, the compound can be a compound of formula (XIX): (XIX) in n is between 3 and 100; k is between 3 and 100; m is 1, 2, 3, or 4; Each g is independently 10, 11, 12, 13, 14, 15, 16, 17 or 18; p is 2, 3, or 4; t is 2, 3, or 4; h is 1, 2, 3, or 4; q is zero or 1; R1 and R2 are independently chosen from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH and -CH2OPO(OH)2, wherein any one of the alkyl hydrogens can be replaced by a halogen, and wherein R1 and R2 are not both H; When q = 1, R3 is -NH2 or -OH; When q = 0, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid. Or a pharmaceutically acceptable salt, solvate, or prodrug.

[0201] In some embodiments, any compound disclosed herein that contains polyethylene glycol (PEG) (including compounds of any one of formulas (I)-(XIX)) may contain a substituted PEG form of PEG.

[0202] In some embodiments, the substituted PEG is represented by the partial formula BI: in n is between 3 and 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; R3 is H, -NH2, or -OH, where R3 is H when q is zero, and R3 is -NH2 or -OH when q is 1; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid.

[0203] In some embodiments, the substituted PEG is represented by the partial formula B-II: B-II in p is 2, 3, or 4; n is between 3 and 100; m is 1, 2, 3, or 4; t is 2, 3, or 4; k is between 3 and 100; h is 1, 2, 3, or 4; q is zero or 1; When q is 1, R3 is -NH2 or -OH; When q is zero, R3 is H; L is empty or consists of 1 to 10 units, each of which is a native α-amino acid or derived from a native α-amino acid, and has the following formula: Where R4 is H; and R5 is the side chain, or the second hydrogen atom of the amino acid.

[0204] In some embodiments of the substituted PEG of formula BI or B-II, q is 1.

[0205] In some embodiments of the substituted PEG of formula BI or B-II, n can be from 10 to 14, such as 11, or from 24 to 30, such as 27.

[0206] In some embodiments of the substituted PEG of formula BI or B-II, m is from 1 to 3, such as 2.

[0207] In some embodiments of the substituted PEG of formula BI or B-II, R3 is -NH2 when q is 1.

[0208] In some embodiments of substituted PEG of formula BI or B-II, L is a natural α-amino acid residue.

[0209] The compounds described herein can exist and be isolated in optically active and racemic forms. As will be understood by those skilled in the art, this invention is intended to cover any racemic, optically active, or stereoisomeric forms, or mixtures thereof, of the compounds of this invention having the useful properties described herein. Methods for preparing these forms (e.g., resolution of racemic mixtures by recrystallization, synthesis from optically active starting materials, chiral synthesis, or separation by chiral chromatography) are well known in the art. In some embodiments, the composition may comprise an enantiomerically or diastereomeric enriched form of the compound. For example, the compound may have at least about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or greater than 99% enantiomer excess (ee) or diastereomeric excess (de). In some embodiments, the compound may be enriched at at least about 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or greater than 99% at any stereocenter of the compound.

[0210] In any respect, the compound may contain a chiral center surrounding the following chiral centers (shown at *): , The chiral center is in the R configuration. This form of the compound can also be referred to as an R-Pam2 analog diastereomer of the compound of the present invention as described herein. The compound can be described as follows: .

[0211] In any respect, the compound may contain a chiral center (shown at *) in the 2,3-bis(palmitoyloxy)propyl moiety of Pam2Cys: , The chiral center is in the R configuration. This form of the compound can also be referred to as the R-Pam2 diastereomer of the compound of the present invention as described herein. The compound can be described as follows: .

[0212] In any respect, the compound may contain a chiral center surrounding the following chiral centers (shown at *): , The chiral center is in the S configuration. This form of the compound can also be referred to as an S-Pam2 analog diastereomer of the compound of the present invention as described herein. The compound can be described as follows: .

[0213] In any respect, the compound contains a chiral center (shown at *) in the 2,3-bis(palmitoyloxy)propyl moiety of Pam2Cys: The chiral center is in the S configuration. This form of the compound can also be referred to as the S-Pam2 diastereomer of the compound of the present invention as described herein. The compound can be described as follows: In any respect, the compound contains a chiral center surrounding the following chiral center (shown at *): , The chiral center is in the L configuration. This form of the compound can also be referred to as an L-Cys analog diastereomer of the Pam2Cys compound described herein. The compound can be described as: .

[0214] In any respect, the compound contains a chiral center (shown at *) in the cysteine ​​residues of Pam2Cys: The chiral center is in the L configuration. This form of the compound can also be referred to as the L-Cys diastereomer of the Pam2Cys compound described herein. The compound can be described as follows: Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0215] In any respect, the compound contains a chiral center in part A1 surrounding the following chiral center (shown at *): , The chiral center is in the D configuration. This form of the compound can also be referred to as the D-Cys analogue diastereomer of the Pam2Cys compound described herein. The compound can be described as: .

[0216] Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0217] In any respect, the compound contains a chiral center (shown at *) in the cysteine ​​residues of Pam2Cys: The chiral center is in the D configuration. This form of the compound can also be referred to as the D-Cys diastereomer of the Pam2Cys compound described herein. The compound can be described as follows: .

[0218] Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0219] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at a chiral center at the carbon atom in portion A2 (shown at *): , The chiral center is in the R configuration. In some embodiments, this stereoisomer of the compound can be described as follows: Among them, L1, L2, Z1, Z2, R x R y R 11 R 12 R13 R 14 R 15 R 16 R 17 R 18 R 19 b, v, and z are as defined for compounds of formula (I), and w is 1. Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0220] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at a chiral center at the carbon atom in portion A2 (shown at *): , The chiral center is in the S configuration. In some embodiments, part A of this stereoisomer of the compound can be described as: Among them, L1, L2, Z1, Z2, R x R y R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 b, v, w, and z are as defined for compounds of formula (I). Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0221] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at a chiral center at a carbon atom in portion A2 (shown at **): The chiral center is in the L configuration. This type of compound can also be referred to as an L-Cys analog stereoisomer of the compound of the present invention. In some embodiments, this stereoisomer of the compound can be described as follows: Among them, L1, L2, Z1, Z2, R x R y R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R19 b, v, w, and z are as defined for compounds of formula (I). Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0222] In any aspect or embodiment of the invention, the compounds of the invention may be provided in a chiral form enriched at a chiral center at a carbon atom in portion A2 (shown at **): The chiral center is in the D configuration. This type of compound can also be referred to as a D-Cys analog stereoisomer of the compound of the present invention. In some embodiments, part A of this stereoisomer of the compound can be described as: , Among them, L1, L2, Z1, Z2, R x R y R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 R 19 b, v, and z are as defined for compounds of formula (I), and w is 1. Other stereocenters in these compounds may be racemic or independently rich in R or S configurations.

[0223] In any respect, the compound contains a chiral center in the Y portion of the compound (shown at *): The chiral center is in the L-configuration. This form of compound can also be referred to as the LY diastereomer of the compound of the present invention described herein.

[0224] In any respect, the compound contains a chiral center in the Y portion of the compound (shown at *): The chiral center is in the D-configuration. This form of compound can also be... In any respect, a composition comprising a compound of the present invention (including any one of formulas (I)-(XIX)) or a pharmaceutically acceptable salt, solvate or prodrug and a pharmaceutically acceptable carrier, diluent or excipient may be used in the methods or uses of the present invention.

[0225] In some embodiments, the compound as described herein is an R diastereomer surrounding the chiral center of the 2,3-bis(palmitoyloxy)propyl moiety of the compound.

[0226] In some embodiments, the compound as described herein is an S diastereomer of the chiral center surrounding the 2,3-bis(palmitoyloxy)propyl moiety of the compound.

[0227] In any respect, the compositions described herein comprise the following compound: the compound is an R diastereomer surrounding the chiral center of the 2,3-bis(palmitoyloxy)propyl moiety of the compound.

[0228] In any respect, the composition comprises a compound that is an S diastereomer of the chiral center surrounding the 2,3-bis(palmitoyloxy)propyl moiety of the compound.

[0229] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are R diastereomers surrounding the chiral center of the 2,3-bis(palmitoyloxy)propyl moiety of the compound.

[0230] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are S diastereomers surrounding the chiral center of the 2,3-bis(palmitoyloxy)propyl moiety (e.g., part A1) of the compound.

[0231] In any respect, the compounds described herein are L diastereomers of the chiral center of the cysteine ​​analog residues surrounding a portion of the Pam2Cys analog compound (e.g., portion Y).

[0232] In any respect, the compounds described herein are L diastereomers of the chiral center of the cysteine ​​residues surrounding the Pam2Cys partial compounds (e.g., the Y partial compounds).

[0233] In any respect, the compounds described herein are D diastereomers of the chiral center of the cysteine ​​analog residues surrounding a portion of the Pam2Cys analog compound (e.g., portion Y).

[0234] In any respect, the compounds described herein are D diastereomers of the chiral center of the cysteine ​​residues surrounding the Pam2Cys moiety (e.g., part Y) of the compound.

[0235] In any respect, the compositions described herein comprise a compound that is an L diastereomer of a chiral center of a cysteine ​​analog residue surrounding a Pam2Cys analog moiety (e.g., part Y) of the compound.

[0236] In any respect, the compositions described herein comprise a compound that is an L diastereomer of a chiral center of a cysteine ​​residue surrounding the Pam2Cys moiety (e.g., part Y) of the compound.

[0237] In any respect, the compositions described herein comprise a compound that is a D diastereomer of a chiral center of a cysteine ​​analog residue surrounding a Pam2Cys analog moiety (e.g., part Y) of the compound.

[0238] In any respect, the compositions described herein comprise a compound that is a D diastereomer of a chiral center of a cysteine ​​residue surrounding the Pam2Cys moiety (e.g., part Y) of the compound.

[0239] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are L diastereomers of chiral centers of cysteine ​​analog residues surrounding the Pam2Cys analog moiety of the compound.

[0240] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are L diastereomers of the chiral center of the cysteine ​​residue surrounding the Pam2Cys moiety of the compound.

[0241] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are D diastereomers of the chiral center of the cysteine ​​analog residues surrounding the Pam2Cys analog moiety of the compound.

[0242] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are D diastereomers of the chiral center of the cysteine ​​residue surrounding the Pam2Cys moiety of the compound.

[0243] In any respect, the compounds of the present invention as described herein are L diastereomers surrounding the chiral center of the Y moiety.

[0244] In any respect, the compounds described herein are D diastereomers surrounding the chiral center of the Y moiety.

[0245] In any respect, the compositions described herein comprise the following compound: the compound is an L diastereomer surrounding the chiral center of the Y moiety.

[0246] In any respect, the compositions described herein comprise a D diastereomer of the chiral center surrounding the Y moiety.

[0247] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are L diastereomers surrounding the chiral center of the Y moiety.

[0248] In any respect, 1%, 5%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, 95%, 96%, 97%, 98%, 99%, or more than 99% of the compound present in the composition are D diastereomers surrounding the chiral center of the Y moiety.

[0249] The compounds of formula (I)-(XIX) described herein exhibit remarkable stability in solution. This solution stability can be observed by storing solutions of the compounds under ambient storage conditions (e.g., at 25°C) or under accelerated degradation stability conditions (e.g., at 40°C) for at least about 14 days.

[0250] In any respect, any of the compounds described herein may be administered in the form of a pharmaceutically acceptable salt.

[0251] The term “pharmaceutical acceptable” can be used to describe any pharmaceutically acceptable salt, hydrate, or prodrug, or any other compound that, when administered to a subject, can (directly or indirectly) provide a compound of the present invention as described herein, or a pharmaceutically acceptable salt, prodrug, or ester thereof, or an active metabolite or residue thereof.

[0252] Suitable pharmaceutically acceptable salts may include, but are not limited to, salts of pharmaceutically acceptable inorganic acids such as hydrochloric acid, sulfuric acid, phosphoric acid, nitric acid, carbonic acid, boric acid, aminosulfonic acid, and hydrobromic acid; or salts of pharmaceutically acceptable organic acids such as acetic acid, propionic acid, butyric acid, tartaric acid, maleic acid, hydroxymaleic acid, fumaric acid, malic acid, citric acid, lactic acid, mucoic acid, gluconic acid, benzoic acid, succinic acid, oxalic acid, phenylacetic acid, methanesulfonic acid, toluenesulfonic acid, benzenesulfonic acid, salicylic acid, sulfanilic acid, aspartic acid, glutamic acid, edemaic acid, stearic acid, palmitic acid, oleic acid, lauric acid, pantothenic acid, tannic acid, ascorbic acid, and valeric acid.

[0253] Alkali salts may include, but are not limited to, those formed with pharmaceutically acceptable cations such as sodium, potassium, lithium, calcium, magnesium, zinc, ammonium, alkylammonium (such as salts formed from triethylamine), alkoxyammonium (such as those formed from ethanolamine), and salts formed from ethylenediamine, choline, or amino acids such as arginine, lysine, or histidine. General information regarding the types of pharmaceutically acceptable salts and their formation is known to those skilled in the art, and is as generally stated in texts such as " Handbook of Pharmaceutical salts [Handbook of Medicinal Salts], PH Stahl, CG Wermuth, 1st ed., 2002, Wiley-VCH.

[0254] When the compounds are solids, those skilled in the art will understand that the compounds, reagents, and salts of the present invention can exist in different crystalline or polymorphic forms, all of which are within the scope of the present invention and the specified chemical formulas.

[0255] The term "polymorph" includes any crystalline form of the compounds of the present invention as described herein, such as anhydrous form, aqueous form, solvate form, and mixed solvate form.

[0256] Where applicable, the compounds of the present invention described herein are intended to cover both solvated and non-solvated forms. Therefore, the compounds of the present invention described herein include compounds having the structures shown, including hydrated or solvated forms as well as non-hydrated and non-solvated forms.

[0257] As used herein, the term "solvent" refers to a complex of variable stoichiometry formed by a solute (in this invention, the compound of the invention described herein or a pharmaceutically acceptable salt, prodrug, or ester thereof) and a solvent. Such solvents used for the purposes of this invention must not interfere with the biological activity of the solute. Examples of suitable solvents include, but are not limited to, water, methanol, ethanol, and acetic acid. Preferably, the solvent used is a pharmaceutically acceptable solvent. Examples of suitable pharmaceutically acceptable solvents include, but are not limited to, water, ethanol, and acetic acid. Most preferably, the solvent used is water.

[0258] Basic nitrogen-containing groups can be quaternized with reagents such as: lower alkyl halides, such as methyl, ethyl, propyl and butyl chlorides, bromides and iodides; dialkyl sulfates such as dimethyl sulfate and diethyl sulfate; etc.

[0259] The compounds described herein also include isotopic variants, such as those using deuterium instead of hydrogen.

[0260] A "prodrug" is a compound that may not fully meet the structural requirements of the compounds provided herein, but is modified in vivo upon administration to a subject or patient to produce the compounds of the present invention as described herein. For example, a prodrug can be an acylated derivative of a compound as provided herein. Prodrugs include compounds in which a hydroxyl, carboxyl, amino, or thiol group is bonded to any of the following groups, which, upon administration to a mammalian subject, cleaves to form a free hydroxyl, carboxyl, amino, or thiol group, respectively. Examples of prodrugs include, but are not limited to, acetate (salts), formate (salts), phosphate (salts), and benzoate (salts) derivatives of alcohol and amine functional groups in the compounds provided herein. Prodrugs of the compounds provided herein can be prepared in such a manner that the functional groups present in the compound are modified such that the modification cleaves in vivo to produce the parent compound.

[0261] Prodrugs comprise compounds in which an amino acid residue, or a polypeptide chain of two or more (e.g., two, three, or four) amino acid residues is covalently linked to the free amino and amide groups of any compound of formula (I)-(XIX). The amino acid residues include the 20 naturally occurring amino acids typically represented by three-letter symbols, and also include 4-hydroxyproline, hydroxylysine, demosine, isodemosine, 3-methylhistidine, norvlin, β-alanine, γ-aminobutyric acid, citrulline, homocysteine, homoserine, ornithine, and methionine sulfone. Prodrugs also comprise compounds in which carbonates, carbamates, amides, and alkyl esters are covalently bonded to the aforementioned substituents of the compounds described herein, including compounds of formula (I)-(XIX) or other structures as described herein.

[0262] The common chemical terms used in the formulas in this article have their usual meaning.

[0263] The term "aliphatic group" is intended to include saturated and unsaturated, non-aromatic, straight-chain, branched, acyclic, and cyclic hydrocarbons. Those skilled in the art will understand that aliphatic groups include, for example, alkyl, alkenyl, alkynyl, cycloalkyl, and cycloalkenyl groups, and their hybrids, such as (cycloalkyl)alkyl, (cycloalkenyl)alkyl, and (cycloalkyl)alkenyl. In various embodiments, the aliphatic group comprises 1 to 12, 1 to 8, 1 to 6, or 1 to 4 carbon atoms. In some embodiments, the aliphatic group comprises 5 to 21, 9 to 21, or 11 to 21 carbon atoms, such as 11, 13, 15, 17, or 19 carbon atoms. In some embodiments, the aliphatic group is saturated.

[0264] The term "heteroaliphatic group" is intended to include an aliphatic group in which one or more chain and / or ring carbon atoms are independently replaced by heteroatoms, preferably selected from oxygen, nitrogen, and sulfur. In some embodiments, the heteroaliphatic group is saturated. Examples of heteroaliphatic groups include straight-chain or branched heteroalkyl, heteroalkenyl, and heteroynyl groups.

[0265] The term "alkyl" is intended to include saturated straight-chain and branched hydrocarbon groups. In some embodiments, the alkyl group has 1 to 12, 1 to 10, 1 to 8, 1 to 6, or 1 to 4 carbon atoms. In some embodiments, the alkyl group has 5 to 21, 9 to 21, or 11 to 21 carbon atoms, such as 11, 13, 15, 17, or 19 carbon atoms. Examples of straight-chain alkyl groups include, but are not limited to, methyl, ethyl, n-propyl, n-butyl, n-pentyl, n-hexyl, n-heptyl, and n-octyl. Examples of branched alkyl groups include, but are not limited to, isopropyl, isobutyl, sec-butyl, tert-butyl, neopentyl, isopentyl, and 2,2-dimethylpropyl.

[0266] The term "alkenyl" is intended to include straight-chain and branched alkyl groups having at least one double bond between two carbon atoms. In some embodiments, the alkenyl group has 2 to 12, 2 to 10, 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, the alkenyl group has 5 to 21, 9 to 21, or 11 to 21 carbon atoms, such as 11, 13, 15, 17, or 19 carbon atoms. In some embodiments, the alkenyl group has one, two, or three carbon-carbon double bonds. Examples of alkenyl groups include, but are not limited to, vinyl, allyl, -CH=CH(CH3), -CH=C(CH3)2, -C(CH3)=CH2, and -C(CH3)=CH(CH3).

[0267] The term "alkynyl" is intended to include straight-chain and branched alkyl groups having at least one triple bond between two carbon atoms. In some embodiments, the alkynyl group has 2 to 12, 2 to 10, 2 to 8, 2 to 6, or 2 to 4 carbon atoms. In some embodiments, the alkynyl group has one, two, or three carbon-carbon triple bonds. Examples include, but are not limited to, -C=CH, -C=CH3, -CH2C=CH3, and -C=CH2CH(CH2CH3)2.

[0268] The term "heteroalkyl" is intended to include alkyl groups in which one or more carbon atoms of the chain are replaced by heteroatoms, preferably selected from the group consisting of oxygen, nitrogen, and sulfur. In some embodiments, the heteroalkyl group is saturated. Heteroalkyl groups include, for example, polyethylene glycol groups and polyethylene glycol ether groups.

[0269] The term "cycloalkyl" is intended to include monocyclic, bicyclic, or tricyclic alkyl groups. In some embodiments, one or more rings of a cycloalkyl group have 3 to 12, 3 to 10, 3 to 8, 3 to 6, or 3 to 5 carbon atoms. In some embodiments, a cycloalkyl group has 5 or 6 ring carbon atoms. Examples of monocyclic cycloalkyl groups include, but are not limited to, cyclopropyl, cyclobutyl, cyclopentyl, cyclohexyl, cycloheptyl, and cyclooctyl. In some embodiments, a cycloalkyl group has 3 to 8, 3 to 7, 3 to 6, 4 to 6, 3 to 5, or 4 to 5 ring carbon atoms. Bicyclic and tricyclic cyclic systems include bridged, spirocyclic, and fused cycloalkyl cyclic systems. Examples of bicyclic and tricyclic cycloalkyl systems include, but are not limited to, bicyclic [2.1.1]hexyl, bicyclic [2.2.1]heptyl, adamantyl, and decahydronaphthyl.

[0270] The term "cycloalkenyl" is intended to include non-aromatic cycloalkyl groups having at least one double bond between two carbon atoms. In some embodiments, the cycloalkenyl group has one, two, or three double bonds. In some embodiments, one or more rings of the cycloalkenyl group have 4 to 14, 5 to 14, 5 to 10, 5 to 8, or 5 to 6 carbon atoms. In some embodiments, the cycloalkenyl group has 5, 6, 7, or 8 cyclic carbon atoms. Examples of cycloalkenyl groups include cyclohexenyl, cyclopentenyl, cyclohexadienyl, butadienyl, pentadienyl, and hexadienyl.

[0271] The term "aryl" is intended to include cyclic aromatic hydrocarbon groups that do not contain any cyclic heteroatoms. Aryl groups include monocyclic, bicyclic, and tricyclic ring systems. Examples of aryl groups include, but are not limited to, phenyl, azulel, heptadeninl, biphenyl, fluorenyl, phenanthryl, anthracenel, indenel, indanyl, cyclopentadienyl, and naphthyl. In some embodiments, one or more rings of the aryl group have 6 to 14, 6 to 12, or 6 to 10 carbon atoms. In some embodiments, the aryl group is phenyl or naphthyl. Aryl groups include aromatic-aliphatic fused ring systems. Examples include, but are not limited to, indenel and tetrahydronaphthyl.

[0272] The term "heterocyclic group" is intended to include non-aromatic ring systems containing three or more ring atoms, wherein one or more of these ring atoms are heteroatoms. In some embodiments, the heteroatom is nitrogen, oxygen, or sulfur. In some embodiments, the heterocyclic group contains one, two, three, or four heteroatoms. In some embodiments, the heterocyclic group includes monocyclic, bicyclic, and tricyclic rings having 3 to 16, 3 to 14, 3 to 12, 3 to 10, 3 to 8, or 3 to 6 ring atoms. Heterocyclic groups include partially unsaturated and saturated ring systems, such as imidazolinyl and imidazoalkyl groups. Heterocyclic groups include fused and bridged ring systems containing heteroatoms, such as quinine cyclic groups. Heterocyclic groups include, but are not limited to, acridine, aziridine, aziridine, diaziridine, 1,3-dioxane, 1,3-dioxolane, isoxazolyl, morpholinyl, piperazine, piperidinyl, pyranyl, pyrazolyl, pyrrolinyl, pyrrolyl, tetrahydrofuranyl, tetrahydrothiophenyl, thiadiazolyl, and trithiayl.

[0273] The term "heteroaryl" is intended to include aromatic ring systems containing five or more ring atoms, wherein one or more of these ring atoms are heteroatoms. In some embodiments, the heteroatoms are nitrogen, oxygen, or sulfur. In some embodiments, heteroaryl includes monocyclic, bicyclic, and tricyclic ring systems having 5 to 16, 5 to 14, 5 to 12, 5 to 10, 5 to 8, or 5 to 6 ring atoms. Heteroaryl groups include, but are not limited to, pyrroloyl, pyrazolyl, triazolyl, tetraazolyl, oxazolyl, isoxazolyl, thiazolyl, pyridinyl, pyridinyl, pyrazinyl, phenylthio, benzobenzylthio, furanyl, benzofuranyl, indole, azindolyl (pyrrolopyridinyl), inazolyl, benzimidazolyl, pyrazolopyridinyl, triazolopyridinyl, benzotriazolyl, benzooxazolyl, benzothiazolyl, imidazopyridinyl, isoxazolopyridinylxanthinyl, guaninyl, quinolinyl, isoquinolinyl, tetrahydroquinolinyl, quinoxalinyl, and quinazolinyl. Heteroaryl groups include fused ring systems in which all rings are aromatic rings, such as indole, and fused ring systems in which only one ring is an aromatic ring, such as 2,3-dihydroindole.

[0274] The term "halogen" is intended to include F, Cl, Br, and I.

[0275] The term "heteroatom" is intended to include oxygen, nitrogen, sulfur, or phosphorus. In some embodiments, heteroatoms are selected from the group consisting of oxygen, nitrogen, and sulfur.

[0276] As used herein, the term "substituted" is intended to mean that one or more hydrogen atoms in the indicated group are replaced by one or more independently selected suitable substituents, provided that the substitution does not exceed the normal valence state of each atom attached to one or more substituents, and that the substitution produces a stable compound. In some embodiments, the optional substituents in the compounds described herein include, but are not limited to, halogens, CN, NO2, OH, NH2, NHR. 100 NR 100 R 200 C 1-6 Haloalkyl, C 1-6 Haloalkoxy, C(O)NH2, C(O)NHR 100 C(O)NR 100 R 200 SO2R 100 OR 100 SR 100 S(O)R 100 C(O)R 100 and C 1-6 Aliphatic groups; of which R 100 and R 200 Each is C independently 1-6 Aliphatic groups, such as C 1-6 alkyl.

[0277] When referring to protecting groups (PG), those skilled in the art will readily understand what type of protecting group would be appropriate.

[0278] As used in this text, the term "amine protecting group" is intended to refer to a group that can be readily removed to provide an NH2 group in the amine group and protects the amine group from unwanted reactions during the synthetic procedure. Such protecting groups are described in Protective Groups in Organic Synthesis by TW Greene et al. (John Wiley & Sons, 1999) and Amino Acid-Protecting Groups by Fernando Albericio (with Albert Isidro-Llobet and Mercedes Alvarez), Chemical Reviews, 2009 (109) 2455-2504. Examples include, but are not limited to, acyl and acyloxy groups, such as acetyl, chloroacetyl, trichloroacetyl chloride, o-nitrophenylacetyl, o-nitrophenoxy-acetyl, trifluoroacetyl, acetoacetyl, 4-chlorobutyryl chloride, isobutyryl, picolinoyl, aminohexanoyl, benzoyl, methoxy-carbonyl, 9-fluorenylmethoxycarbonyl, 2,2,2-trifluoroethoxycarbonyl, 2-trimethylsilylethoxy-carbonyl, tert-butyloxycarbonyl, benzyloxycarbonyl, p-nitrobenzyloxycarbonyl, 2,4-dichlorobenzyloxycarbonyl, etc. Other examples include Cbz (carboxybenzyl), Nosyl ((o-nitrophenylsulfonyl or p-nitrophenylsulfonyl), Bpoc (2-(4-biphenyl)isopropoxycarbonyl), and Dde (1-(4,4-dimethyl-2,6-dioxane)ethyl). In some embodiments, the amine protecting groups used for the purposes described herein include (but are not limited to) tert-butoxycarbonyl (t-Boc) and 9H-fluorene-9-ylmethoxycarbonyl (Fmoc).

[0279] As used in this text, the term "carboxyl protecting group" is intended to mean a group that can be readily removed to provide an OH group in the carboxyl group and protects the carboxyl group from unwanted reactions during the synthetic procedure. Such protecting groups are described in *Protective Groups in Organic Synthesis* by TW Greene et al. (John Willie & Son, 1999) and in *Amino Acid-Protecting Groups* by Fernando Albericio (with Albert Isidro-Llobet and Mercedes Alvarez), *Chemical Reviews*, 2009 (109) 2455-2504. Examples include, but are not limited to, alkyl and silyl groups, such as methyl, ethyl, tert-butyl, methoxymethyl, 2,2,2-trichloroethyl, benzyl, diphenylmethyl, trimethylsilyl, and tert-butyldimethylsilyl, etc.

[0280] As used in this text, the term "formamide protecting group" is intended to refer to a group that can be readily removed to provide an NH2 group in the formamide group and protects the formamide group from unwanted reactions during the synthetic procedure. Such protecting groups are described in *Protective Groups in Organic Synthesis* by TW Greene et al. (John Willie & Son, 1999) and in *Amino Acid-Protecting Groups* by Fernando Albericio (with Albert Isidro-Llobet and Mercedes Alvarez), *Chemical Reviews*, 2009 (109) 2455-2504. Examples include, but are not limited to, 9-oxazanthyl (Xan), triphenylmethyl (Trt), methyltriphenyl (Mtt), cyclopropyl dimethyl progenityl (Cpd), and dimethylcyclopropylmethyl (Dmcp).

[0281] The term "ester" refers to a carboxylic acid group in which the hydrogen atom of the hydroxyl group has been replaced by a saturated straight-chain (i.e., linear) or branched hydrocarbon group. Specific examples of alkyl groups are methyl, ethyl, propyl, isopropyl, n-butyl, isobutyl, sec-butyl, tert-butyl, n-pentyl, isopentyl, n-hexyl, and 2,2-dimethylbutyl. Alkyl groups can be C1-C6 alkyl groups. As used herein, terms defining the limits of a length range, such as "from 1 to 5," mean any integer from 1 to 5, i.e., 1, 2, 3, 4, and 5. In other words, any range defined by two explicitly mentioned integers is intended to include and disclose any integer defining the limit and any integers included in the range. Alkyl groups can be branched alkyl groups.

[0282] As used in this article, "Ser" refers to the amino acid serine, and "Cys" refers to the amino acid cysteine.

[0283] As used herein, “PEG” refers to the polymer compound polyethylene glycol. Unless otherwise defined, references to “PEG” include ethylene oxide polymers of any length. References to PEG also include substituted PEGs. In some embodiments, substituted PEGs may be defined by formula BI or B-II as described herein.

[0284] As used in this article, the term "and / or" means "and" or "or", or both.

[0285] The term "one or more" preceding a noun considers both singular and plural forms, or both forms simultaneously.

[0286] References to the numerical ranges disclosed herein (e.g., 1 to 10) are intended to also include references to all rational numbers within that range (e.g., 1, 1.1, 2, 3, 3.9, 4, 5, 6, 6.5, 7, 8, 9, and 10), as well as any range of rational numbers within that range (e.g., 2 to 8, 1.5 to 5.5, and 3.1 to 4.7), and therefore, all subranges of the entire ranges explicitly disclosed herein are hereby explicitly disclosed. These are merely examples of specific intent, and all possible combinations of numerical values ​​between the listed minimum and maximum values ​​should be considered as expressly stated in a similar manner in this application.

[0287] Immune system stimulants In one aspect of the invention, a method for treating, preventing, or minimizing cancer progression is provided, comprising administering a TLR2 agonist and an immunostimulant.

[0288] Those skilled in the art should understand that the terms “immunostimulant,” “immunostimulating,” or “stimulate / induce immune response,” as used herein, and their grammatical equivalents, mean to induce, increase, enhance, or otherwise provide a beneficial effect on the immune response.

[0289] "Immune stimulation," "immunostimulatory," or "stimulated / induced immune response" refers to a direct or indirect response of immune system cells or components to any treatment described herein. Such responses can be measured in any manner known in the art, including activation, proliferation, or differentiation of immune system cells (B cells, T cells, dendritic cells, APCs, macrophages, NK cells, NKT cells, etc.), upregulation or downregulation of serum markers, cytokines, interferons, IgM, and IgG release, and mixed cell infiltration in various organs.

[0290] Immunostimulants can include specific and non-specific immunostimulants. Specific immunostimulants provide antigen specificity in an immune response, such as vaccines or any antigen. Non-specific immunostimulants enhance the immune response to other antigens or stimulate components of the immune system in a manner independent of antigen specificity, such as adjuvants and non-specific immunostimulants.

[0291] These immunostimulants can assist the immune system in any one or more different ways, including by stimulating antigen-presenting cells, T cells, or innate cells to trigger and enhance the immune system; by modulating suppressive pathways to reduce immunosuppression in the tumor environment; and / or by enhancing adaptive or innate immune responses. Therefore, stimulation of the immune system can be the stimulation of innate or acquired immune system cells. Immunostimulants can inhibit immunosuppression induced by cancer cells or antigen-presenting cells.

[0292] As used in this article, immunostimulants can be molecules that do not directly stimulate the immune system but can repolarize immune cells or suppress immunosuppression. Alternatively, immunostimulants can directly stimulate or activate the immune system.

[0293] Innate immunity refers to those immune responses that occur rapidly following cancer infection or development. They begin without prior sensitization to the pathogen or malignant cells, lack antigen specificity, and are directly mediated by phagocytes such as macrophages, cytotoxic cells such as natural killer (NK) cells, and antigen-presenting cells such as dendritic cells (DCs), and indirectly mediated by cytokines produced by these cells. Adaptive immunity, or cell-mediated immunity, refers to those responses that require a period of development after initial infection or cancer development and involve the education of immune cells to produce highly specific, highly effective, and long-lasting responses. This is mediated by cytotoxic T lymphocytes (CTLs), helper T lymphocytes, and antibody-producing B lymphocytes. Following these lines of thought, adaptive immune responses are classified as cellular responses (those mediated by CTLs) or humoral responses (antibody-mediated responses), with helper T lymphocytes promoting both. In summary, rapid innate immune responses play a role in controlling the early spread of disease and promoting the development of adaptive immune responses, while highly effective, specific, and long-lasting adaptive responses are used to clear disease and prevent recurrence.

[0294] This invention contemplates the use of immunostimulants in combination with any TLR2 agonists described herein for the treatment, prevention, or maximal reduction of cancer progression. Suitable immunostimulants that may be used according to the methods described herein include: - Cellular immunotherapy (cytotoxic cell immunotherapy or adoptive cell therapy). - Oncolytic virus; - Cancer vaccines; - T-cell binding agents; and - Bispecific T-cell conjugate.

[0295] Specific examples of suitable immunostimulants include Blincyto (blinatumomab), Oncotice (BCG [strain Tice] vaccine), BCG (BCG [strain Rivm] vaccine), ImmuCyst, Pacis (BCG [strain Montréal] vaccine), Provenge (sipuleucel-T), DCVax-L (DCVax-L), Oncorine (human adenovirus type 5 [recombinant]), and Imlygic (talimogene laherparepvec).

[0296] Suitable immunostimulants for use in the methods of the present invention also include activators of co-stimulatory molecules or inhibitors of immune checkpoint molecules. Thus, the immunostimulant can be an immune checkpoint inhibitor, a co-stimulatory molecule agonist, or an immune activator. Examples of checkpoint inhibitors that can be used in the present invention are described herein. Inhibition of the inhibitory molecule can be achieved by inhibition at the DNA, RNA, or protein level. In embodiments, inhibitory nucleic acids (e.g., dsRNA, siRNA, or shRNA) can be used to inhibit the expression of the inhibitory molecule. In other embodiments, the inhibitor of the inhibitory signal is a peptide, such as a soluble ligand, or an antibody or antigen-binding fragment thereof that binds to the inhibitory molecule.

[0297] The co-stimulatory molecule can be any of the following co-stimulatory molecules selected from one or more agonists of OX40, CD2, CD27, CDS, ICAM-1, LFA-1, (CD11a / CD18), ICOS (CD278), 4-lBB (CD137), GITR, CD30, CD40, BAFFR, HVEM, CD7, LIGHT, NKG2C, SLAMF7, NKp80, CD160, B7-H3, or CD83 ligands. Agonists of any of these co-stimulatory molecules are also considered immunostimulants.

[0298] Checkpoint inhibitors "Checkpoint inhibitors" inactivate proteins in the inhibitory checkpoint pathway of the immune response.

[0299] In the case of cancer, checkpoint inhibitors modulate the immune system by blocking proteins that prevent the immune system from attacking cancer cells. Specifically, they control how evasive cancer cells and T cells interact, allowing T cells to recognize tumor cells and generate an appropriate immune response against them. Non-limiting examples of checkpoint inhibitors that can be used according to the methods described herein include inhibitors targeting PD-1 (programmed cell death protein 1), CTLA-4 (cytotoxic T-lymphocyte-associated protein 4), and PD-L1 (programmed death ligand 1). Those skilled in the art will understand that CTLA-4 and PD-1 are found on T cells, and PD-L1 is expressed on cancer cells. Other non-limiting examples include PD-L2, TIM3, LAG3, CEACAM (e.g., CEACAM-1, CEACAM-3 and / or CEACAM-5), VISTA, BTLA, TIGIT, LAIR1, CD160, 2B4, CD80, CD86, B7-H3 (CD276), B7-H4 (VTCN1), HVEM (TNFRSF14 or CD107), KIR, A2aR, MHC class I, MHC class II, GAL9, adenosine, and TGF-β.

[0300] Other immune checkpoints include indoleamine 2,3-dioxygenase (IDO) and CSF-R1. Inhibitors of these proteins are also considered to be immune checkpoint inhibitors for use in this invention.

[0301] Programmed death-1 (PD-1) refers to an immunosuppressive receptor belonging to the CD28 family. PD-1 is primarily expressed on activated T cells in vivo and binds to two ligands, PD-L1 and PD-L2. As used herein, the term "PD-1" includes human PD-1 (hPD-1), variants, isotypes, and species homologs of hPD-1, as well as analogs having at least one epitope shared with hPD-1. The complete hPD-1 sequence can be found in GenBank accession number U64863.

[0302] Upon binding of PD-1 to programmed cell death ligand 1 (PD-L1), the immune response is shut down to prevent T cells from destroying or killing cells. In the case of cancer, cancer cells can be covered by the PD-L1 protein to disguise themselves as healthy cells, thereby evading the immune response. Programmed death ligand-1 (PD-L1) is one of two cell surface glycoprotein ligands of PD-1 (the other being PD-L2), which downregulates T cell activation and cytokine secretion upon binding to PD-1. As used herein, the term “PD-L1” includes human PD-L1 (HPD-L1), variants, isotypes, and species homologs of hPD-L1, as well as analogs having at least one epitope shared with hPD-L1. The complete hPD-L1 sequence can be found under GenBank accession number Q9NZQ7.

[0303] Cytotoxic T-lymphocyte antigen-4 (CTLA-4) is an immunosuppressive receptor belonging to the CD28 family. CTLA-4 is expressed exclusively on T cells in vivo and binds to two ligands, CD80 and CD86 (also known as B7-1 and B7-2, respectively). As used herein, the term "CTLA-4" includes human CTLA-4 (hCTLA-4), variants, isotypes, and species homologs of hCTLA-4, as well as analogs having at least one epitope shared with hCTLA-4. The complete hCTLA-4 sequence can be found in GenBank accession number AAB59385.

[0304] Any checkpoint inhibitor described herein may be administered in the form of an antibody. An “antibody” (Ab) should include, but is not limited to, a glycoprotein immunoglobulin or its antigen-binding portion that specifically binds to an antigen and comprises at least two heavy (H) chains and two light (L) chains interconnected by disulfide bonds. Each H chain contains a heavy chain variable region (abbreviated VH) and a heavy chain constant region. The heavy chain constant region contains three constant domains CH1, CH2, and CH3. Each light chain contains a light chain variable region (abbreviated VL) and a light chain constant region. The light chain constant region contains one constant domain CL. The VH and VL regions may be further subdivided into hypervariable regions called complementarity-determining regions (CDRs) and scattered with more conserved regions called framework regions (FRs). Each VH and VL contains three CDRs and four FRs, arranged in the following order from the amino terminus to the carboxyl terminus: FR1, CDR1, FR2, CDR2, FR3, CDR3, FR4. The variable regions of the heavy and light chains contain binding domains that interact with the antigen. The constant region of Ab mediates the binding of immunoglobulins to host tissues or factors, including various cells of the immune system (such as effector cells) and the first component (Clq) of the classical complement system.

[0305] The term "antibody" includes, for example, monoclonal and polyclonal Abs, chimeric and humanized Abs, human or non-human Abs, fully synthetic Abs, and single-chain Abs. Non-human Abs can be humanized by recombinant methods to reduce their immunogenicity in humans. Unless otherwise specified, the term "antibody" also includes antigen-binding fragments or portions of any of the above-described immunoglobulins, including monovalent and bivalent fragments or portions, as well as single-chain Abs.

[0306] "Isolated antibody" refers to an Ab that is substantially free of other Abs with different antigen specificities (e.g., an isolated Ab that specifically binds to PD-1 is substantially free of Abs that specifically bind to antigens other than PD-1). However, isolated Abs that specifically bind to PD-1 may be cross-reactive with other antigens, such as PD-1 molecules from different species. Furthermore, isolated Abs may be substantially free of other cellular material and / or chemicals. The term "monoclonal antibody" (mAb) refers to an Ab molecule formulation that is not naturally occurring and has a single molecular composition; that is, an Ab molecule whose primary sequence is substantially identical and exhibits a single binding specificity and affinity for a specific epitope. mAbs are examples of isolated Abs. mAbs can be produced by hybridoma, recombinant organisms, transgenic organisms, or other techniques known to those skilled in the art.

[0307] A "human" antibody (huMAb) is an Ab having a variable region, wherein both the framework region and the CDR region are derived from human germline immunoglobulin sequences. Furthermore, if the Ab contains a constant region, that constant region is also derived from a human germline immunoglobulin sequence. The human Abs of this invention may include amino acid residues not encoded by human germline immunoglobulin sequences (e.g., mutations introduced by random or site-specific mutagenesis in vitro or by somatic mutations in vivo). However, as used herein, the term "human antibody" is not intended to include Abs in which a CDR sequence derived from another mammalian species, such as a mouse, has been grafted onto a human framework sequence. The terms "human" Ab and "fully human Ab" are used synonymously.

[0308] "Humanized antibody" refers to an Ab in which some, most, or all of the amino acids outside the CDR domain of a non-human Ab are replaced by corresponding amino acids derived from human immunoglobulins. In one embodiment of the humanized form of an Ab, some, most, or all of the amino acids outside the CDR domain have been replaced by amino acids derived from human immunoglobulins, while some, most, or all of the amino acids within one or more CDR regions remain unchanged. Minor additions, deletions, insertions, substitutions, or modifications of amino acids are permitted, as long as they do not eliminate the Ab's ability to bind to a specific antigen. "Humanized" Abs retain antigen specificity similar to the original Ab.

[0309] "Chimeric antibody" refers to an antibody whose variable region originates from one species and whose constant region originates from another species, such as an antibody whose variable region originates from a mouse antibody and whose constant region originates from a human antibody.

[0310] An "anti-antigen" Ab refers to an Ab that specifically binds to an antigen. For example, an anti-PD-1 Ab specifically binds to PD-1, and an anti-CTLA-4 Ab specifically binds to CTLA-4.

[0311] An "antigen-binding portion" (also known as an "antigen-binding fragment") of an Ab refers to one or more fragments of an Ab that retain the ability to specifically bind antigens bound by the entire Ab.

[0312] Examples of immune checkpoint inhibitors and antibody inhibitors targeting those checkpoints include anti-CTLA-4 (e.g., ipilimumab, tremelimumab, KAHR-102), anti-TIM3 (e.g., F38-2E2, ENUM005), anti-LAG3 (e.g., BMS-986016, IMP701, IMP321, C9B7W), anti-KIR (e.g., lirilumab, IPH2101, IPH4102), and anti-PD-1 (e.g., nivolumab, pitilizumab, pembrolizumab, BMS-936559, atezolizumab). b) Lambrolizumab, MK-3475, AMP-224, AMP-514, STI-A1110, TSR-042, anti-PD-L1 (e.g., KY-1003 (EP20120194977), MCLA-145, atezolizumab, BMS-936559, MEDI-4736, MSB0010718C, AUR-012, STI-A1010, PCT / US2001 / 020964, MPDL3280A, AMP-224, dafiloribumab (polyethylene glycol, CDP-7657), MEDI-4920), anti-CD73 (e.g., AR-42 (OSU-HDAC42, HDAC-42, AR42, AR... 42. OSU-HDAC 42. OSU-HDAC-42, NSC D736012, HDAC-42, HDAC 42, HDAC42, NSCD736012, NSC-D736012), MEDI-9447), anti-B7-H3 (e.g., MGA271, DS-5573a, 8H9), anti-CD47 (e.g., CC-90002, TTI-621, VLST-007), anti-BTLA, anti-VISTA, anti-A2aR, anti-B7-1, anti-B7-H4, anti-CD52 (e.g., alemtuzumab), anti-IL-10, anti-IL-35, anti-TGF-β (e.g., Fresolumimab), anti-CSF1R (e.g., FPA008), anti-NKG2A (e.g., monalizumab), anti-MICA (e.g., IPH43), and anti-CD39.

[0313] Anti-PD-1 antibody and anti-PD-L1 antibody Examples of suitable PD-1 inhibitors that may be used according to the present invention include Keytruda (pembrolizumab), Opdivo (nivolumab), AAGEN 2034, BGB-A317, BI-754091, CBT-501 (genolimzumab), MEDI0680, MGA012, PDR001, PF-06801591, REGN2810 (SAR439684), and TSR-042, or those disclosed in U.S. Patent No. 8,008,449. Other anti-PD-1 mAbs have been described, for example, in U.S. Patent Nos. 6,808,710, 7,488,802, 8,168,757, and 8,354,509 and PCT Publication No. WO 2012 / 145493.

[0314] Nivolumab (also known as “Opdivo®”; formerly known as 5C4, BMS-936558, MDX-1106, or ONO4538) is a fully human IgG4 (S228P) PD-1 immune checkpoint inhibitor Ab that selectively blocks the interaction with PD-1 ligands (PD-L1 and PD-L2), thereby blocking the downregulation of anti-tumor T cell function (US Patent No. 8,008,449).

[0315] Pembrolizumab (also known as Keytruda®, lambrolizumab, and MK-3475) is a humanized monoclonal IgG4 antibody targeting the human cell surface receptor PD-1 (programmed death-1). Pembrolizumab is described, for example, in U.S. Patent Nos. 8,354,509 and 8,900,587. Pembrolizumab has been approved by the FDA for the treatment of relapsed or refractory melanoma.

[0316] Other suitable PD-1 inhibitors include Libtayo (cimiplimab), Blincyto, Dostarlimab, Spartalizumab, Cetrelimab, Pidilizumab, and BI-754091.

[0317] The anti-PD-1 antibody used in the disclosed methods or compositions is an immunosuppressive antibody that binds to PD-1 with high specificity and affinity, blocks the binding of PD-L1 and / or PD-L2, and inhibits the PD-1 signaling pathway. In any composition or method disclosed herein, the anti-PD-1 antibody comprises an antigen-binding portion or fragment that binds to the PD-1 receptor and exhibits similar functional properties to the intact antibody in inhibiting ligand binding and upregulating the immune system.

[0318] In some embodiments, the anti-PD-1 antibody used in these methods can be replaced by another PD-1 or anti-PD-L1 antagonist. For example, since anti-PD-L1 antibodies prevent the interaction between PD-1 and PD-L1, thereby exerting a similar effect to the PD-1 signaling pathway, anti-PD-L1 antibodies can be used instead of anti-PD-1 antibodies in the methods disclosed herein. In any embodiment, suitable PD-L1 inhibitors include Imfinzi (durvalumab or MEDI4736), Tecentriq (atezolizumab or MPDL3280A), Bavencio (avelumab; MSB0010718C), MS-936559 (12A4 or MDX-1105), and CX-072.

[0319] Anti-CTLA-4 antibody The anti-CTLA-4 antibody of this invention binds to human CTLA-4, thereby disrupting the interaction between CTLA-4 and the human B7 receptor. It should be understood that since the interaction between CTLA-4 and B7 transduces signals leading to the inactivation of T cells carrying the CTLA-4 receptor, disruption of this interaction effectively induces, enhances, or prolongs the activation of such T cells, thereby inducing, enhancing, or prolonging the immune response.

[0320] Suitable CTLA-4 inhibitors that may be used according to the present invention include Yervoy (ipilimumab), trimemumab, and those disclosed in AGEN 1884 or U.S. Patent Nos. 6,984,720 and 7,605,238. Ipilimumab is a fully human IgG1 monoclonal antibody that blocks the binding of CTLA-4 to its B7 ligand, thereby stimulating T cell activation. Trimemumab is a human IgG2 monoclonal anti-CTLA-4 antibody. Another is Blincyto (bonatumab), a bispecific CD19-directed CD3 T-cell conjugate.

[0321] Application and dosage In one embodiment of the invention, a therapeutically effective amount of a TLR2 agonist and a checkpoint inhibitor is administered to the subject.

[0322] Administration refers to the physical introduction of a composition containing a therapeutic agent into a subject using any of the various methods and delivery systems known to those skilled in the art, including those described herein. Pharmaceutical compositions can be formulated from compounds of the invention as described herein for any suitable route of administration. Typically, in addition to a therapeutic agent (e.g., a TLR2 agonist and / or an immunostimulant), a pharmaceutical composition also contains pharmaceutically acceptable excipients, carriers, and / or diluents. Examples of suitable components included in pharmaceutical compositions are described in Martindale - The Extra Pharmacopoeia (Pharmaceutical Press, London, 1993) and Martin (ed.), Remington's Pharmaceutical Sciences.

[0323] Suitable routes of administration for carrying out the defined methods include oral, intravenous, intramuscular, local, subcutaneous, intraperitoneal, spinal, or other parenteral administration routes, such as by injection or infusion. As used herein, the phrase “parenteral administration” refers to a mode of administration other than enteral and local administration, typically by injection, and includes, but is not limited to, intravenous, intramuscular, intraarterial, intrasheath, intralymphatic, intralesional, intracapsular, intracardiac, intradermal, intraperitoneal, tracheal, subcutaneous, subepidermal, intra-articular, subcapsular, subarachnoid, intraspinal, epidural, and intrasternal injections and infusions, as well as intracorporeal electroporation. Administration may also be performed, for example, once, multiple times, and / or over one or more extended time periods.

[0324] Any of the compositions described herein can be formulated for application to the respiratory tract, in other words, via the respiratory route. When considering application to all or part of the respiratory tract, those skilled in the art should understand that this includes intranasal or inhalation administration, particularly for administration to the lungs. Compositions as described herein can be formulated for intranasal administration, including dry powder, spray, mist, or aerosol.

[0325] Formulations suitable for application, for example as nasal sprays or nasal drops (where the carrier is liquid), include aqueous or oily solutions of the active ingredient. Alternatively, the composition may be a dry powder and may be applied only to the respiratory tract as defined herein.

[0326] The selection of a suitable carrier depends on the specific type of application intended. For administration via the respiratory tract (e.g., the nasal mucosa), the compound can be formulated as a buffered or non-buffered solution (e.g., water or isotonic saline), or as a suspension, for intranasal administration as drops or sprays. Preferably, such solutions or suspensions are isotonic relative to nasal secretions and have approximately the same pH, for example, from about pH 4.0 to about pH 7.4 or from pH 6.0 to pH 7.0. The buffer should be physiologically compatible and include, for example, phosphate buffers. For example, representative nasal decongestants are described as buffered to a pH of about 6.2 (Remington's, Id., p. 1445). Of course, those skilled in the art can readily determine the appropriate saline content and pH of a harmless aqueous carrier for nasal and / or respiratory tract administration.

[0327] Other ingredients may also be included, such as preservatives, colorants, lubricating or viscous mineral or vegetable oils, fragrances, natural or synthetic plant extracts such as aromatic oils, and humectants and viscosity enhancers such as glycerin, to provide the formulation with additional viscosity, moisturizing properties, and a pleasant texture and odor. For nasal administration of the solution or suspension according to the invention, various devices in the art can be used to produce drops, liquids, and sprays. For example, the TLR2 agonists and / or immunostimulants or compositions described herein can be administered into the nasal passage via a simple dropper (or pipette) comprising a glass, plastic, or metal dispensing tube from which contents are dispensed dropwise by air pressure provided by a manually driven pump (e.g., a flexible rubber bulb) attached to one end.

[0328] Tears from the eyes drain from the orbit into the nasal passages; therefore, if desired, suitable pharmaceutically acceptable ophthalmic solutions can be readily provided by a person of ordinary skill as a carrier for delivering the compounds or compositions described herein, and can be applied to the orbit in the form of eye drops for both ocular and intranasal administration.

[0329] In one embodiment, a pre-measured unit dose dispenser is prepared comprising a dropper or spray device for delivery of a solution or suspension in the form of drops or sprays, containing one or more doses of the drug to be administered. The invention also includes a cartridge containing one or more dehydrated doses of a compound, along with any desired salts and / or buffers, preservatives, colorants, etc., which can be prepared into a solution or suspension by adding an appropriate amount of water. The water may be sterile or non-sterile, but sterile water is generally preferred.

[0330] The phrase “therapeutic effective amount” or “effective amount” generally refers to the amount of the TLR2 agonist and / or checkpoint inhibitor, its pharmaceutically acceptable salt, polymorph, or prodrug of the present invention that has the effect of: (i) treating a particular disease, condition, or disorder; (ii) reducing, improving, or eliminating one or more symptoms of a particular disease, condition, or disorder; or (iii) delaying the onset of one or more symptoms of a particular disease, condition, or disorder as described herein. Undesirable effects (e.g., side effects) may occur alongside the desired therapeutic effect; therefore, practitioners will weigh the potential benefits against the potential risks when determining what constitutes an appropriate “effective amount.”

[0331] For example, in treating tumors, a therapeutically effective amount of the compounds or compositions described herein can inhibit tumor growth by at least about 10%, at least about 20%, at least about 30%, at least about 40%, at least about 50%, at least about 60%, at least about 70%, at least about 80%, or at least about 90% or more, relative to an untreated subject. Alternatively, the treatments described herein can cause complete regression of the tumor mass. In other embodiments of the invention, tumor regression can be observed and persist for at least about 10 days, at least about 20 days, at least about 30 days, at least about 40 days, at least about 50 days, or at least about 60 days, at least about 70 days, at least about 80 days, at least about 90 days, at least about 100 days, or longer.

[0332] Therapeutic effective amounts of a drug may also include “prophylactic” or “preventive effective amounts,” which are any amounts of a TLR2 agonist and / or checkpoint inhibitor administered to a subject at risk of developing cancer (e.g., a subject with precancerous lesions) or a subject experiencing cancer recurrence that inhibit cancer development or recurrence. In some embodiments, preventive effective amounts completely prevent cancer development or recurrence. “Inhibit” or “prevent” cancer development or recurrence means reducing the likelihood of cancer development or recurrence, or completely preventing cancer development or recurrence.

[0333] The exact amount of therapeutically effective dose required will vary from subject to subject, depending on the subject's species, age and general condition, method of administration, etc. Therefore, it may be impossible to specify an exact therapeutically effective dose. However, those skilled in the art can determine the appropriate therapeutically effective dose for any individual situation using routine experiments. In one aspect, the dose administered to the subject is any therapeutically effective dose that results in a reduction of cancer-related symptoms as a result of any of the following: a reduction in the number of cancer cells; a reduction in tumor size; inhibition (i.e., to some extent slowing and preferably stopping) of cancer cell infiltration into surrounding organs; inhibition (i.e., to some extent slowing and preferably stopping) of tumor metastasis; inhibition of tumor growth to some extent; or relief to some extent from one or more cancer-related symptoms. Additionally or alternatively, a therapeutically effective dose may result in an increase in the subject's survival rate.

[0334] In some embodiments, the therapeutically effective amount of a TLR2 agonist for a human subject is in the range of about 250 nmol / kg body weight / dose to 0.005 nmol / kg body weight / dose. Preferably, this range is about 250 nmol / kg body weight / dose to 0.05 nmol / kg body weight / dose. In some embodiments, the body weight / dose range is about 250 nmol / kg to 0.1 nmol / kg, about 50 nmol / kg to 0.1 nmol / kg, about 5 nmol / kg to 0.1 nmol / kg, about 2.5 nmol / kg to 0.25 nmol / kg, or about 0.5 nmol / kg to 0.1 nmol / kg body weight / dose. In some embodiments, this amount is or is about 250 nmol, 50 nmol, 5 nmol, 2.5 nmol, 0.5 nmol, 0.25 nmol, 0.1 nmol, or 0.05 nmol / kg body weight / dose of the compound. Dosing regimens can be adjusted to suit emergency situations and can be further adjusted to obtain the optimal therapeutic dose.

[0335] Typically, therapeutically effective doses are formulated to contain at least about 0.1% up to about 50% or higher (by weight), and all combinations and sub-combinations within that range. Compositions may be formulated to contain one or more compounds or their pharmaceutically acceptable salts, polymorphs, or prodrugs at concentrations from about 0.1% to less than about 50% (e.g., about 49%, 48%, 47%, 46%, 45%, 44%, 43%, 42%, 41%, or 40%), including concentrations from greater than about 0.1% (e.g., about 0.2%, 0.3%, 0.4%, or 0.5%) to less than about 40% (e.g., about 39%, 38%, 37%, 36%, 35%, 34%, 33%, 32%, 31%, or 30%). Exemplary compositions may contain concentrations from about 0.5% to less than about 30% (e.g., about 29%, 28%, 27%, 26%, 25%, 25%, 24%, 23%, 22%, 21%, or 20%), including concentrations from more than about 0.5% (e.g., about 0.6%, 0.7%, 0.8%, 0.9%, or 1%) to less than about 20% (e.g., about 19%, 18%, 17%, 16%, 15%, 14%, 13%, 12%, 11%, or 10%). Compositions may contain concentrations from more than about 1% (e.g., about 2%) to less than about 10% (e.g., about 9% or 8%), including concentrations from more than about 2% (e.g., about 3% or 4%) to less than about 8% (e.g., about 7% or 6%). The active agent may be present, for example, at a concentration of about 5%. In all cases, the amount may be adjusted to compensate for variations in the amount of active ingredient actually delivered to the treated cells or tissues.

[0336] For the administration of checkpoint inhibitors, including PD-1, PD-L1, or CTLA-4 inhibitors, the dosage can range from about 0.01 mg / kg to about 20 mg / kg, about 0.1 mg / kg to about 10 mg / kg, about 0.1 mg / kg to about 5 mg / kg, about 1 mg / kg to about 5 mg / kg, about 2 mg / kg to about 5 mg / kg, about 7.5 mg / kg to about 12.5 mg / kg, or about 0.1 mg / kg to about 30 mg / kg of the subject's body weight. For example, the dosage can be about 0.1 mg / kg, about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, about 5 mg / kg, or about 10 mg / kg of body weight, or about 0.3 mg / kg, about 1 mg / kg, about 2 mg / kg, about 3 mg / kg, or about 5 mg / kg of body weight. Dosing regimens are typically designed to achieve sustained receptor occupancy (RO) exposure based on the typical pharmacokinetic properties of the inhibitor. Exemplary treatment regimens require administration approximately once weekly, approximately once every two weeks, approximately once every three weeks, approximately once every four weeks, approximately once monthly, approximately once every three to six months, or longer. In some embodiments, the checkpoint inhibitor is administered to the subject approximately every two weeks. In other embodiments, the Ab is administered approximately every three weeks. Dosage and timing may be varied during treatment. For example, an anti-PD-1 therapy dosing regimen may include administration of the Ab at the following frequencies: (i) approximately once every two weeks over a period of approximately six weeks; (ii) approximately once every four weeks for approximately six doses, followed by approximately once every three months; (iii) approximately once every three weeks; (iv) approximately 3 mg / kg to approximately 10 mg / kg once, followed by approximately 1 mg / kg every approximately two to three weeks. Given that IgG4 Ab typically has a half-life of 2-3 weeks, the dosing regimen of the anti-PD-1 Ab of the present invention comprises intravenous administration of about 0.3 mg / kg to about 0 mg / kg body weight, 1 mg / kg to 5 mg / kg body weight, or about 1 mg / kg to about 3 mg / kg body weight, wherein the Ab is administered every about 14-21 days in a cycle of up to about 6 weeks or about 12 weeks, until a complete response or confirmed progressive disease.

[0337] In some embodiments, the checkpoint inhibitor and / or TLR2 agonist treatment disclosed herein lasts for at least about 1 month, at least about 2 months, at least about 3 months, at least about 4 months, at least about 5 months, at least about 6 months, at least about 7 months, at least about 8 months, at least about 9 months, at least about 10 months, at least about 11 months, at least about 1 year, at least about 18 months, at least about 24 months, at least about 3 years, at least about 5 years, or at least about 10 years.

[0338] It should be understood that the specific dose level for any particular patient will depend on a variety of factors, including the activity of the specific compound used, age, weight, general health condition, sex, diet, time of administration, route of administration, excretion rate, combination of drugs (i.e., other drugs used to treat the patient), and the severity of the specific disorder being treated.

[0339] The term "treatment" for a subject includes the application or administration of the compounds of this invention to a subject with the aim of delaying, slowing, stabilizing, curing, healing, reducing, alleviating, altering, correcting, reducing the worsening of, alleviating, improving, or affecting a disease or condition, its symptoms, or the risk (or susceptibility) to a disease or condition. The term "treatment" also refers to any indication of successfully treating or improving an injury, lesion, or condition, including any objective or subjective parameter such as elimination; relief; slowing the rate of exacerbation; reducing the severity of the disease; stabilizing, reducing symptoms, or making the injury, lesion, or condition more tolerable for the subject; slowing the rate of degeneration or decline; making the degenerative endpoint less debilitating; or improving the physical or mental health of the subject.

[0340] As used herein, minimizing or preventing cancer progression refers to treating a subject to prevent or delay tumor recurrence or metastasis, or to prevent the growth of an existing tumor. Minimizing or preventing cancer progression includes preventing or delaying cancer recurrence after cancer treatment, or preventing the growth of an existing tumor. Recurrences being prevented include, for example, recurrence in the tumor bed after surgical resection. Alternatively, recurrence includes metastasis of cancer to another part of the body. The terms “preventing recurrence” and “preventing relapse” as used herein are interchangeable.

[0341] This invention also includes methods for preventing the development of cancer in an individual. For example, an individual requiring cancer prevention may be considered at risk of developing cancer but does not yet have detectable cancer. An individual at risk of cancer development may be an individual with a family history of cancer, and / or an individual whose genetic testing or other tests indicate a high risk or high probability of developing cancer. The individual may have cancer stem cells but has not yet been found to have any detectable tumor. It should be understood that methods for preventing cancer development include methods for delaying the onset of cancer in a subject.

[0342] The terms "subject" and "patient" should be understood as interchangeable. While this invention can be used in humans, it is also useful for therapeutic veterinary purposes. This invention can be used with livestock or farm animals such as cattle, sheep, horses, and poultry; pets such as cats and dogs; and zoo animals.

[0343] cancer The term “cancer” should be understood to include benign, precancerous, pretumoral, or non-metastatic tumors or metastatic tumors.

[0344] In some embodiments, the type of cancer to be treated includes those with benign, precancerous, pretumoral, or nonmetastatic tumors. A benign tumor should be understood as one that is not malignant and does not invade nearby tissues or spread to other parts of the body. Similarly, a nonmetastatic cancer should be understood as one that does not invade nearby tissues or spread to other parts of the body. "Precancerous" or "pretumoral" generally refers to a condition or growth that typically precedes or develops into cancer. "Precancerous" growth may have cells characterized by abnormal cell cycle regulation, proliferation, or differentiation, which can be identified by cell cycle markers.

[0345] In one embodiment, the cancer is a secondary cancer or a metastatic tumor. Secondary cancers can be located in any organ or tissue, particularly those with relatively high hemodynamic pressure, such as the lungs, liver, kidneys, pancreas, intestines, and brain. Secondary cancers can be detected in ascites and / or lymph nodes.

[0346] Precancerous, neoplastic, and metastatic cancers are specific examples to which the methods of this invention can be applied. A broad range of examples include breast tumors, colorectal tumors, adenocarcinoma, mesothelioma, bladder tumors, prostate tumors, germ cell tumors, hepatocellular carcinoma / cholangiocarcinoma, neuroendocrine tumors, pituitary tumors, small round cell tumors, squamous cell carcinoma, melanoma, atypical fibroxanthoma, seminoma, non-seminomatous tumor, mesenchymal linedichoma, Sertoli cell tumor, skin tumors, kidney tumors, testicular tumors, brain tumors, ovarian tumors, gastric tumors, oral cavity tumors, bladder tumors, bone tumors, cervical tumors, esophageal tumors, laryngeal tumors, liver tumors, lung tumors, vaginal tumors, and Wilms' tumors.

[0347] Specific examples of cancers include, but are not limited to, adenocarcinoma, adenoma, fibroadenoma, adenolymphoma, adenoma, AIDS-related cancers, acoustic neuroma, acute lymphoblastic leukemia, acute myeloid leukemia, cystic adenoid carcinoma, adrenocortical carcinoma, idiopathic myeloid metaplasia, alopecia areata, alveolar soft tissue sarcoma, ameloblastoma, angiokeratoma, eosinophilic angiolymphoid hyperplasia, hemangioma sclerosis, hemangioma, amine precursor uptake decarboxylation cell tumor, anal cancer, angiosarcoma, aplastic anemia, astrocytoma, ataxia-telangiectasia, and basal cell carcinoma (skin). Bladder cancer, bone cancer, colon cancer, brainstem glioma, brain and CNS tumors, breast cancer, branchial protozoa, CNS tumors, carcinoid tumors, cervical cancer, childhood brain tumors, childhood cancers, childhood leukemia, childhood soft tissue sarcoma, chondrosarcoma, choriocarcinoma, chronic lymphocytic leukemia, chronic myeloid leukemia, colorectal cancer, cutaneous T-cell lymphoma, carcinomas (e.g., Walker's carcinoma, basal cell carcinoma, squamous basal cell carcinoma, Brown-Pearce tumor), ductal carcinoma, Ehrlich tumor, Krebs 2 carcinoma. 2) Merkel cell carcinoma, mucinous carcinoma, non-small cell lung cancer, oat cell carcinoma, papillary carcinoma, scleroderma, bronchiolar carcinoma, bronchial carcinoma, squamous cell carcinoma, and transitional cell carcinoma, carcinosarcoma, cervical dysplasia, phyllodes tumor, cementum tumor, chordoma, pustular cystoma, chondrosarcoma, chondroblastoma, craniopharyngioma, chondroma, cholesteatoma, cylindrica, cystadenocarcinoma, cystadenoma, dermatofibrosarcoma protuberans, and fibroblastic microsatellite carcinoma. Cytoplasmic tumor, ductal carcinoma, dysgerminoma, endocrine carcinoma, endometrial cancer, ependymoma, esophageal cancer, Ewing's sarcoma, extrahepatic bile duct cancer, ocular cancer, melanoma, retinoblastoma, fallopian tube cancer, Fanconi anemia, fibroma, fibrosarcoma, gallbladder cancer, gastric cancer, gastrointestinal cancer, gastrointestinal carcinoid, urogenital cancer, germ cell tumors, gestational trophoblastic disease, glioma, gynecological cancer, giant cell tumor, ganglioma, glioma, vascular tumor Tumors, granulomas, amyotrophic lateral sclerosis (ALS), hematologic malignancies, hairy cell leukemia, head and neck cancer, hepatocellular carcinoma, hereditary breast cancer, histiocytosis, Hodgkin's disease, human papillomavirus (HPV), hydatidiform mole, hypercalcemia, hypopharyngeal cancer, hamartoma, hemangioendothelioma, hemangioma, hemangiopericytoma, angiosarcoma, angiosarcoma, histiocytic diseases, malignant histiocytosis, histiocytoma, hepatocellular carcinoma, hidradenoma, chondrosarcoma, immunoproliferative small cell tumor, opoma, intraocular melanoma, islet cell carcinoma, Kaposi's sarcoma, renal cancer, Langerhans cell histiocytosis, laryngeal cancer, leiomyosarcoma, leukemia, Livroumin syndrome, lip cancer, liposarcoma, liver cancer, lung cancer, lymphedema, lymphoma, Hodgkin's lymphoma, non-Hodgkin's lymphoma, leiomyosarcoma, leukemia (e.g., B-cell, mixed cell, naked cell, T-cell, chronic T-cell, HTLV-II related).Lymphangiosarcoma, acute lymphoblastic, chronic lymphoblastic, mast cell and myeloid cell lymphoma, leukemic sarcoma, Ledich's cell tumor, liposarcoma, leiomyoma, leiomyosarcoma, lymphangioma, lymphangiocytoma, lymphangioma, lymphangiomyoma, lymphangiosarcoma, male breast cancer, malignant rhabdomyosarcoma of the kidney, medulloblastoma, melanoma, Merkel cell carcinoma, mesothelioma, metastatic carcinoma, oral cancer, multiple endocrine tumors, mycosis fungoides, myelodysplastic syndrome, myeloma, myeloproliferative disorders, malignant carcinoid syndrome, carcinoid heart disease, medulloblastoma Meningioma, melanoma, mesenchymal tumor, mesonephroma, mesothelioma, myoblastoma, fibroid, sarcoma, myxoma, myxosarcoma, nasal carcinoma, nasopharyngeal carcinoma, nephroblastoma, neuroblastoma, neurofibromatosis, Niemeh's rupture syndrome, non-melanoma skin cancer, non-small cell lung cancer (NSCLC), schwannoma, neuroblastoma, neuroepithelial tumor, neurofibromatosis, neurofibroma, neuroma, tumors (e.g., bone, breast, digestive system, colorectal, liver), eye cancer, esophageal cancer, oral cancer, oropharyngeal cancer, osteosarcoma, ostomy, ovarian cancer, pancreatic cancer. Paranasal carcinoma, parathyroid carcinoma, parotid gland carcinoma, penile cancer, peripheral neuroectodermal tumors, pituitary carcinoma, polycythemia vera, prostate cancer, osteoma, osteosarcoma, ovarian cancer, papilloma, paraganglioma, paraganglioma non-pheochromocytoma, pineal tumor, plasmacytoma, proto-oncogenes, rare cancers and related diseases, renal cell carcinoma, retinoblastoma, rhabdomyosarcoma, Rosmond-Thomson syndrome, reticuloendothelial hyperplasia, rhabdomyosarcoma, salivary gland carcinoma, sarcoma, schwannoma, Cézare syndrome, skin cancer, small cell lung cancer (SCLC), small intestinal cancer, soft tissue sarcoma. Tumors, spinal cord tumors, squamous cell carcinoma (skin), gastric cancer, synovial sarcoma, sarcomas (e.g., Ewing's experimental sarcoma, Kaposi's sarcoma, and mast cell sarcoma), Seltoli cell carcinoma, synovial sarcoma, testicular cancer, thymic carcinoma, thyroid cancer, transitional cell carcinoma (bladder), transitional cell carcinoma (kidney-pelvis- / ureter), trophoblastic carcinoma, teratoma, theca cell carcinoma, thymoma, trophoblastic carcinoma, urethral cancer, urinary system cancer, uroplakin, uterine sarcoma, uterine cancer, vaginal cancer, vulvar cancer, Waldenström macroglobulinemia, and Wilms' tumor.

[0348] The presence, improvement, treatment, or maximal reduction of cancer progression can be determined by any clinical or biochemically relevant method as described herein or known in the art. A positive response to treatment or maximal reduction of cancer progression can be determined by any method known in the art and may include determining: - The number of cancer cells is reduced; - Tumor size decreased; - Inhibit (i.e., slow down and preferably stop) the infiltration of cancer cells into surrounding organs; - Inhibit (i.e., slow down and preferably stop) tumor metastasis to some extent; -Reducing or completely preventing tumor metastasis after resection of the primary tumor; - It can inhibit tumor growth to a certain extent; - To some extent, alleviate one or more symptoms associated with cancer; and / or -Increased subject survival rate.

[0349] A determination of any of the above can be considered a positive response to the TLR agonists and / or checkpoint inhibitors described herein.

[0350] In contrast, a negative or lack of response to treatment, including any TLR agonist and / or checkpoint inhibitor described herein, can be determined by any method known in the art, and may include determining: - The number of cancer cells did not change or increase; - The tumor size has not changed or increased; - Cancer cells infiltrate surrounding organs without change, persistence, or increase; - Tumor metastasis remains unchanged, persists, or increases; -Despite the removal of the primary tumor, there was no change or increase in tumor metastasis; - Tumor growth showed no change or increase; - One or more cancer-related symptoms have not changed or have increased; and / or - Subject survival rates did not change or decrease.

[0351] In one embodiment of the invention, the subject may have previously received treatment. In one embodiment, the prior treatment is a checkpoint inhibitor, which may be in the form of an inhibitor of PD-1, PD-L1, or CTLA-4. In a preferred embodiment, the checkpoint inhibitor is in the form of an antibody.

[0352] Subjects who have previously received cancer treatment may experience partial or complete remission. In other words, as described above, subjects who have previously received cancer treatment may experience a 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%, or more reduction in measurable parameters of tumor growth, which can be detected through physical examination, radiological studies, or by biomarker levels from blood or urine tests. Alternatively, in the case of complete remission, all detectable disease manifestations completely disappear, leaving the subject without any detectable signs of cancer. Subjects may have substantially undetectable signs of cancer. “Substantially undetectable” cancer generally refers to a situation where treatment has exhausted the size, volume, or other physical measure of the cancer, making it undetectable by relevant standard detection techniques (e.g., in vivo imaging) as a result of treatment.

[0353] The goals or outcomes of treatment with TLR2 agonists and / or checkpoint inhibitors can be to reduce the number of cancer cells; reduce the size of the primary tumor; inhibit (i.e., to some extent slow down and preferably stop) cancer cell infiltration into surrounding organs; inhibit (i.e., to some extent slow down and preferably stop) tumor metastasis; inhibit tumor growth to some extent; and / or alleviate one or more symptoms associated with the disorder to some extent. The efficacy of treatment can be measured by assessing duration of survival, time to disease progression, response rate (RR), duration of response, and / or quality of life.

[0354] In one embodiment, this method is particularly useful for delaying cancer progression. In another embodiment, this method is particularly useful for prolonging subject survival, including overall survival and progression-free survival. It should be understood that overall survival is the length of time a patient survives from the date of cancer diagnosis or the start of treatment. It should be understood that progression-free survival is the length of time a patient has the disease but has not progressed during or after cancer treatment.

[0355] Survival analysis can be performed using techniques well-known in the art, including the Kaplan-Mayer method. The Kaplan-Mayer method estimates a survival function based on survival data. In medical research, it can be used to measure the proportion of patients who survive a certain period after treatment. The survival function plot of the Kaplan-Mayer method is a series of decreasing horizontal steps that, with a sufficiently large sample size, approximate the true survival function of the population. The survival function values ​​are assumed to be constant between consecutive different sampled observations (“clicks”).

[0356] A key advantage of the Kaplan-Mayer curve is that it can account for “censored” data—sample loss before the final outcome is observed (e.g., if a patient drops out of the study). On the graph, small vertical scale markers indicate where patient data has been censored. When no truncation or censoring occurs, the Kaplan-Mayer curve is equivalent to the empirical distribution.

[0357] In one embodiment, the method is particularly useful for providing a complete response to treatment, whereby all signs of cancer disappear in response to the treatment. This does not always mean that the cancer has been cured. In one embodiment, the method is particularly useful for providing a partial response to treatment, whereby the size of one or more tumors or lesions, or the extent of cancer in the body, has decreased in response to the treatment.

[0358] medicine box In another embodiment, a kit or article is provided comprising a TLR2 agonist and / or checkpoint inhibitor as described herein, a pharmaceutically acceptable salt, a diluent or excipient as described above, and / or a pharmaceutical composition. Furthermore, the kit may contain instructions for use in any method or application of the invention as described herein.

[0359] In other embodiments, a medicine box is provided for the above-described treatment and / or preventive applications, the medicine box comprising: - Containers that contain therapeutic compositions in the form of TLR2 agonists and / or checkpoint inhibitors as described herein, or pharmaceutically acceptable salts, diluents, excipients, or pharmaceutical compositions; - Labels or packaging instructions with usage instructions.

[0360] In some embodiments, the cassette may contain one or more other active elements or ingredients for treating cancer.

[0361] A medicine box or "article" may include a container and a label or packaging instructions on or associated with that container. Suitable containers include, for example, bottles, vials, syringes, blister packs, etc. These containers may be formed from a variety of materials such as glass or plastic. The container contains a therapeutic composition that effectively treats a condition and may have a sterile access port (e.g., the container may be an intravenous solution bag or a vial with a stopper that can be punctured by a hypodermic needle). The label or packaging instructions indicate that the therapeutic composition is intended for the treatment of the selected condition. In one embodiment, the label or packaging instructions include instructions for use and indicate that the therapeutic or preventative composition may be used to treat the cancer described herein.

[0362] The medicine box may include (a) a therapeutic or preventative composition; and (b) a second container containing a second active element or ingredient. The medicine box in this embodiment of the invention may further include a packaging instruction indicating that the composition and other active elements may be used to treat cancer or prevent the progression of the cancer described herein.

[0363] As used herein, the following compounds are described in the table below, and their specific structures are shown elsewhere herein and are covered in any method or use of the invention. The compounds of the present invention can be prepared by techniques known in the art. For example, compounds of the present invention comprising any one of formulas (I)-(XIX) containing the A1 moiety can be prepared by the techniques described in WO 2019 / 119067, the entire contents of which are hereby incorporated herein by reference.

[0364] The compounds of the present invention comprising any one of formulas (I)-(XIX) containing the A2 portion can be provided by coupling the following two: compounds of formulas A2-I: (A2-I) Where L1, L2, Z1, Z2, v, b, w, z, R x R y R 11 R 12 R 13 R 14 R 15 R 16 R 17 R 18 X has the meaning as defined herein for any compound of the invention, and R 19 It is an amino protecting group Compounds with formula (YB-I): (YB-I) in Y' is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; B' stands for polyethylene glycol (PEG); and It is a solid-phase carrier resin.

[0365] In some embodiments, B' comprises a substituted PEG of formula BI. In these embodiments, a solid-phase reaction of the following sequence may be employed: a) Optionally, use Fmoc chemical components to couple 1 to 10 α-amino acids or compounds derived from natural α-amino acids (constituting L) to a solid resin. b) If L exists, then PG-NH-(CH2) p -O-(CH2CH2O) n -(CH2) m -COOH is coupled to a solid resin or a substituted resin, where PG represents an amino protecting group compatible with the Fmoc chemical composition. c) Remove PG; d) Coupling of PG-NH-CR 13 R 14 -COOH, where PG' represents an amino protecting group compatible with the chemical composition of Fmoc; e) Remove PG'; f) Acids with coupling (AI) structure; g) Optional removal of R 19 And optionally, acylation and / or alkylation are performed to introduce R. 18 and / or R 19 ;as well as h) Removal of the compound from the solid support In some embodiments, B' comprises a substituted PEG according to formula (B-II) and can be subjected to a solid-phase reaction in the following sequence: a) Optionally, use Fmoc chemical components to couple 1 to 10 α-amino acids or compounds derived from natural α-amino acids (constituting L) to a solid-phase resin. b) If L exists, then PG-NH-(CH2)t -O-(CH2CH2O) k -(CH2) h -COOH is coupled to a solid resin or a substituted resin, where PG represents an amino protecting group compatible with the Fmoc chemical composition. c) Remove PG; d) Coupling of PG'-NH-(CH2) p -O-(CH2CH2O) n -(CH2) m -COOH, where PG' represents an amino protecting group compatible with the chemical composition of Fmoc; e) Remove PG'; f) Coupling PG''-NH-CR 13 R 14 -COOH, where PG'' represents an amino protecting group compatible with the chemical composition of Fmoc; g) Remove PG''; h) Acids with coupling (AI); i) Optional removal of R 19 And optionally, acylation and / or alkylation are performed to incorporate R. 18 and / or R 19 ;as well as j) Remove the compound from the solid resin.

[0366] It should be understood that the exact order of events may differ from the order outlined, and additional steps may be added where necessary and synthetically advantageous, such as oxidizing cysteine ​​sulfoxide to sulfoxide or sulfone.

[0367] In some embodiments, the compound of formula A2-I is provided in the form of the compound of formula A2-II: A2-II L1, L2, X, v, w, and R 18 As defined for compounds of the above formula AI, Z1 and Z2 are independently selected from -NHC(O)-, -C(O)NH-, -OC(O)-, -C(O)O-, -NHC(O)O- and -OC(O)O-.

[0368] Compound A2-II can be prepared by the synthesis shown in Scheme 1.

[0369] Scheme 1 describes the synthesis of an example of a compound of formula A2-II, wherein X is S, L1-Z1 is -OC(O)E-(CH2) g-CH3, where E is -O- or -NH-, and g is 10, 11, 12, 13, 14, 15, 16, 17, or 18; L2-Z2 is -OC(O)E-(CH2) g -CH3, where E is -O- or -NH-, and g is 10, 11, 12, 13, 14, 15, 16, 17, or 18; and R 19 It is PG3, which is an amino protecting group.

[0370] Option 1 The reaction of a protected enol of formula (V') (where PG is a suitable protecting group (e.g., silyl such as TBDMS)) forms an epoxide of formula (VI'). It should be understood that epoxide formation can proceed to obtain a racemic product or enantiomer-enriched material. If a racemic or scalemic mixture of enantiomers is produced, preparative chiral chromatography is used to separate the enantiomers (if necessary).

[0371] Under reducing conditions, the epoxide of formula (VI') is reacted with a suitable protected cystine analogue, such as... N -(((9H-fluorene-9-yl)methoxy)carbonyl)-S-(((R)-2-((((9H-fluorene-9-yl)methoxy)carbonyl)amino)-3-(tert-butoxy)-3-oxopropyl)thio)-D-cysteine ​​tert-butyl ester reacts to give the alcohol of formula (VII'), wherein PG2 is tert-butyl ester and PG3 is Fmoc. It should be understood that the alcohol of formula (VII') can consist of more than one stereoisomer, and in the presence of multiple stereoisomers, they can be separated as needed by chiral preparative chromatography.

[0372] The alcohol of formula (VII') can be acylated using a suitable reagent to obtain the carbonyl-containing adduct of formula (VIII'). Where an ester is desired, the acyl chloride can be reacted in the presence of a suitable base and solvent; where a carbamate is desired, the isocyanate can be reacted in the presence of a suitable base and solvent; and where a carbonate is desired, the chloroformate can be reacted in the presence of a suitable base and solvent. The carbonyl-containing adduct of formula (VIII') can then be deprotected using a suitable reagent to expose the carboxylic acid of formula (IX'), for example, in the case that PG2 is tert-butyl, the tert-butyl group can preferably be removed using trifluoroacetic acid.

[0373] The acid of formula (IX') can then be used as a reagent in solid-phase synthesis to add groups of formulas Y and B.

[0374] The compounds of the present invention comprising any one of formulas (I)-(XIX) containing the A2 portion (where z is 1, w is 1, and b is 0) can be provided by preparing a resin-bound peptide having the following formula: in Y' is R1 and R2 are independently selected from the group consisting of: H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=O)NH2, -CH2CH2C(=O)OH, and -CH2CH2C(=O)OR8, wherein any one of the alkyl hydrogens can be replaced by a halogen; R8 is selected from the group consisting of H and straight-chain or branched C1-C6 alkyl groups; B' stands for polyethylene glycol (PEG); PG s It is an H or sulfur protecting group, such as tert-butyl; and It is a solid-phase carrier resin.

[0375] Following optional sulfur deprotection, the resin-bound peptide can be reacted with a 1,2-epoxy-alkanol having the following formula: Where R x R y v has a meaning given for part A2, such as as defined in equation (I). To provide alkylated thiols of formula S-1: S-1 Y' and B' have the meanings given above, and v has the meaning given for part A2, or its sulfone or sulfoxide.

[0376] The diol moiety of the resin-bound compound S-1 can be further reacted (e.g., by diol functionalization with palmitic acid groups or lauryl carbamate groups, etc.) to provide the compound of the present invention.

[0377] Example Examples 1-6 below describe various properties of the compounds of the present invention. The compounds tested in these examples were prepared as described in WO2019 / 119067 or in Synthesis Example 5.

[0378] Example 1: The effect of combination therapy on tumor growth, survival, and metastasis Blocking inhibitory immune cell receptors (also known as “immune checkpoints”) has revolutionized cancer treatment. Despite incredibly high response rates, most patients do not respond to immune checkpoint blockade (ICB) or develop resistance. Little is known about their underlying mechanisms, and there remains an unmet need to develop new or improved immunotherapies. Here, we set out to investigate the role of a synthetic TLR2 agonist, denoted as compound A101 (also referred to as compound 1 in these examples and figures), in combination with anti-PD1 immunotherapy in various cancer and cancer metastasis models.

[0379] Materials and methods mice C57BL / 6 or Balb / c wild-type (WT) mice were purchased from the Walter and Eliza Hall Institute for Medical Research or housed and maintained at the QIMR Berghofer Medical Research Institute. Mice older than 8 weeks were sex-matched to appropriate models. The number of mice treated in each group or the mouse strain for each experiment is shown in the legend. In all studies, no mice were excluded according to pre-established criteria, and randomization was applied immediately prior to treatment in treatment experiments. Experiments were conducted with the approval of the QIMR Berghofer Medical Research Institute Animal Ethics Committee.

[0380] Cell culture Mouse B16F10 (melanoma), MC38 (colon adenocarcinoma), and 4T1.2 (breast cancer) cells were grown in DMEM (Durbeco Modified Eagle Medium) supplemented with 10% fetal bovine serum (Bovogen), 1% glutamine (Gibco), and 1% penicillin / streptomycin (Gibco). B16F10 and 4T1.2 cells were maintained at 37°C and 5% CO2. MC38 cells were maintained at 37°C and 10% CO2. All cell lines were negative for routine mycoplasma testing, but routine cell line identification was not performed.

[0381] Subcutaneous tumor model For primary tumor growth experiments, MC38 (1 × 10⁻⁶) was used. 6 (each) or B16F10 (1 × 10) 5 (Number of cells) were subcutaneously injected into mice at a final volume of 100 μl (day 0). Treatment of mice was initiated as shown in the attached figure or illustration. The vertical diameter of each individual tumor was measured using digital calipers. Tumor size was calculated and expressed as mean ± SEM. When the tumor reached 150 mm...2 At that time, the mice were euthanized.

[0382] 4T1.2 Breast Cancer Metastasis Model 5 × 10 4 Four T1.2 cells were injected at a volume of 50 μl into the fourth mammary fat pad (day 0). On day 12 post-injection, the primary tumor was surgically removed under isoflurane anesthesia. Mice were treated with injections on days 15, 18, and 21 post-injection, as shown in the attached illustration. All mice were sacrificed on day 26 post-injection to assess metastatic burden. For this purpose, grossly visible lung metastases were counted.

[0383] Immune checkpoint blockade assay Immunotherapy was initiated when the transplanted tumor reached a diameter of 3–5 mm. Therapeutic blockade of PD-1 was performed via intraperitoneal injection of 250 μg of rat anti-mouse PD-1 IgG2a (clone RMP1-14; BioXcell) or control rat IgG2a mAb (clone 2A3; BioXcell) in 100 µl PBS solution. Dosage amounts and exact treatment regimens are described in the accompanying figures and / or illustrations.

[0384] Intratumoral injection of compound 1 / compound A101 Mice were also treated with compound 1 when they were first injected with anti-PD1 or control IgG. For this purpose, 100 µl of a saline solution of compound 1 was injected into the tumor. Dosage and treatment regimen are described in the figures and / or legend.

[0385] Intravenous injection of compound 1 / compound A101 Mice were also treated with compound 1 when they were first injected with anti-PD1 or control IgG. For this purpose, 10 μg of compound 1 in 200 μl of saline solution was injected into the lateral tail vein. The treatment regimen is described in the accompanying figures and / or legend.

[0386] result MC38 cells were subcutaneously injected into C57BL / 6 WT mice, and treatment was administered as shown in the figure once the tumor was palpable. Figure 1 A). Combining anti-PD1 immunotherapy with compound 1 significantly improved the survival rate of mice carrying MC38 tumors. Figure 1 B).

[0387] In the next experiment, the B16F10 melanoma cell line, which exhibited poor immunogenicity and resistance to immunotherapy, was injected into C57BL / 6 WT mice. Similarly, when the tumor was palpable, the mice were randomly divided into four groups and treated as shown in the figure. Figure 2Surprisingly, mice carrying B16F10 treated with compound 1 and anti-PD1 showed improved survival rates. Figure 2 (B). Importantly, anti-PD1 monotherapy does not affect survival.

[0388] In summary, this demonstrates that compound 1, when administered topically into the tumor microenvironment, is effectively combined with anti-PD1 immunotherapy in both highly immunogenic and low-immunogenic preclinical cancer models.

[0389] While local injection is possible, it presents challenges in routine clinical practice. Furthermore, many cancerous entities cannot be injected intratumorally. Therefore, the therapeutic efficacy of systemically administered compound 1 in combination with anti-PD1 immunotherapy was explored. For this purpose, MC38 cells were subcutaneously injected into C57BL / 6 WT mice. Once the tumor was palpable, mice were intravenously (iv) injected with 10 μg of compound 1 or a control, and intraperitoneally (ip) injected with anti-PD1 or control IgG. Figure 3 (A). Compared with anti-PD1 monotherapy, the combination immunotherapy of compound 1 and anti-PD1 is superior. Figure 3 B).

[0390] These data indicate that compound 1 enhanced the efficacy of anti-PD1 immunotherapy in MC38 and B16F10 tumor models. Since metastatic dissemination of tumor cells is a leading cause of death in cancer patients, the efficacy of the combination immunotherapy in a spontaneous metastasis model was next evaluated. For this purpose, Balb / c WT mice were injected into the fourth mammary fat pad of a highly aggressive 4T1.2 breast cancer cell line. Similar to typical clinical methods, the primary tumors of the subjects were surgically removed, and the mice were treated with compound 1 alone or in combination with anti-PD1 immunotherapy. Figure 4 (A). All mice were sacrificed 14 days post-surgery, and the number of lung metastases was assessed. Primary 4T1.2 tumors and disseminated cancer cells were highly resistant to anti-PD1 immunotherapy. Systemic administration of 10 μg of compound 1 resulted in a reduction in the number of lung metastases. Importantly, the combination immunotherapy significantly reduced the metastatic burden in mice (A). Figure 4 (B), without affecting weight loss (data not shown).

[0391] In summary, we demonstrate that compound 1 (a highly specific TLR2 agonist) enhances the efficacy of anti-PD1 immunotherapy, providing a fundamental principle for using such combination therapies to treat cancers with diverse etiologies and pathogenesis. These results also demonstrate the efficacy of TLR2 agonist and checkpoint inhibitor combination therapy in cancers previously resistant to checkpoint inhibitor therapy.

[0392] Example 2. Effects of combination therapy on tumor growth and survival in the EMT6.5 model.

[0393] 100,000 EMT6.5 cells were implanted into the fourth inguinal mammary gland of Balb / c mice (n = 48), and the formed tumor was monitored by caliper measurement. When the tumor was ~100 mm... 3 Mice were randomly assigned to one of six treatment groups (n = 8 mice / group). To test additional TLR2 agonists, compound A108 was administered intravenously to tumor-bearing mice in three divided doses, three days apart. On the same day, a checkpoint inhibitor was administered intraperitoneally. The treatment regimen containing compound A108 induced mild but reversible weight loss (data not shown). Data showed that the combination of compound A108 and anti-PDL-1 enhanced the effect on tumor growth (…). Figure 5 (A, C). The combination of compound A108 with anti-PL-1 also showed improved survival ( Figure 5 B).

[0394] These data indicate that repeated administration of systemic compound A108 was well tolerated in Balb / c mice carrying EMT6.5 tumors, and that repeated administration of compound A108 and anti-PDL-1 could statistically enhance the efficacy of different types of checkpoint inhibitors.

[0395] Example 3. Effects of combination therapy on tumor growth and survival in mice carrying MC38.

[0396] Compound A108 was also tested in the MC-38 model in the presence of a checkpoint inhibitor. Highly immunogenic MC38 colon cancer cells were subcutaneously injected into C57BL / 6 WT mice (n = 10-14). Once the tumor was palpable (~3-5 mm diameter), mice were randomly assigned to four groups to receive three intratumoral injections of the drug, anti-PD-1 (200 μg, ip), 25 μg of compound A108 in 100 μl saline solution, or a combination of compound A108 and anti-PD-1. This study showed that the combination therapy improved survival rates in mice. Figure 6 ).

[0397] The above studies were further explored by examining the effect of combination therapy on large tumors in the MC38 mouse model. These studies aimed to understand whether repeated administration over two weeks could slow tumor growth. Mice were administered the drug every two days via intraperitoneal (10 μg dose) for 13 days. Mice were culled upon reaching the humane endpoint or one week after the final dose (up to day 25, whichever came first). Humane endpoints included a weight loss of >20% relative to body weight on day 1 of treatment (or a weight loss of >15% over 3 consecutive days) and an individual mouse tumor volume >3000 mm². 3Or the average tumor volume of a group is > 2000 mm 3 (All mice in this group were excluded.) This study demonstrates that combination therapy can slow tumor growth via an intraperitoneal route during the treatment phase. Figure 7 ).

[0398] These data suggest that the combination of compound A108 and anti-PD1 immunotherapy can weaken tumor growth and prolong the survival of mice carrying MC38 colon cancer.

[0399] Example 4. Compound A108 exhibits antitumor activity in the WEHI164 model.

[0400] This study aimed to determine whether a low dose of compound A108 (2.5 μg) combined with anti-CTLA4, anti-PDL1, or anti-PD-1 could inhibit tumor growth. Data from this experiment demonstrated that when administered in the presence of compound A108, anti-PDL1 (… Figure 8 A), anti-CTLA-4 ( Figure 8 B) and anti-PD-1 ( Figure 8 C) has a greater role.

[0401] These data indicate that, in the WEHI-164 sarcoma cancer model, compound A108 increased the response rate to anti-CTLA4, anti-PDL1, and anti-PD-1 checkpoint therapies.

[0402] Example 5. Representative synthesis, characterization, and TLR2 activity of the selected compounds of this invention: A107, A108, A115, A116, A117, A118, A203, A204, A215, A216, A220, and A224 Synthesis of compounds A107 (x = 11) and A108 (x = 27) Fmoc S-2,3-bis(palmitoyloxypropyl)-cysteine ​​(S-Fmoc-Dpc-OH) was purchased from Bachem Inc.

[0403] Coupling of S-Fmoc-Dpc-OH with resin-bound peptides Fmoc-Dpc-OH (100 mg, 0.24 mmol) was activated for 5 minutes at 0°C with HOBt (36 mg, 0.24 mmol) and N,N'-diisopropylcarbodiimide (DICI; 37 μL, 0.24 mmol) in DCM and DMF (1:1, v / v, 3 mL). The mixture was then added to a solution containing Boc-Cys-Ser(tBu)CH2CH2O-(PEG). 11-CH2CH2C(O)Gly resin or Boc-Cys-Ser(tBu)-CH2CH2O-(PEG) 27 The solution was placed in a container with -CH2CH2C(O)Gly resin (0.25 mmol / g, 0.25 g = 0.0625 mmol). After shaking for 2 hours, the solution was filtered off through a sintered glass funnel (porosity 3), and the resin was washed with DCM and DMF (3 × 30 mL each). The completion of the reaction was monitored using a trinitrobenzenesulfonic acid (TNBSA) test. If necessary, a bicoupling was performed.

[0404] Cleavage of peptides from a solid support: Reagent B (93% TFA, 5% water, and 2% triisopropylsilane) was kept for two hours. The peptide did not precipitate in cold ether. Most of the TFA had to be removed, and the residue was then dissolved in 50% acetonitrile and purified immediately or freeze-dried.

[0405] Synthesis of compounds A115 and A116.

[0406] The synthesis of compounds A115 (x = 11) and A116 (x = 27) was carried out as described in Scheme 2. (R)-glycidyl was coupled to a thiol group linked to a cysteine ​​residue of a peptide resin via alkylation; to 250 mg of saturated Boc-Cys-Ser(tBu)CH2CH2O-(PEG) in DMF. 11 -CH2CH2C(O)Gly resin or Boc-Cys-Ser(tBu)CH2CH2O-PEG 27250 µl of R-(+)-glycidyl (MW = 74.08, d = 1.1, 250 µl = 3.71 mmol, equivalent to a 60-fold excess of free thiol groups on the peptide resin) and 25 µl of diisopropylethylamine (DIPEA, MW = 129.2, d = 0.74, 25 µl = 0.14 mmol) were added to CH2CH2C(O)Gly resin (0.25 mmol / g, 0.25 g = 0.0625 mmol). The reaction mixture was incubated in a 50°C water bath for 2 hours, and then the solid support was thoroughly washed with DMF. 100 µl of ethyl methyl sulfide (W = 76.16, d = 0.842, 100 µl = 1.10 mmol) was added to 250 mg of peptide resin after glycidification and washing with toluene, followed by 105 µl of tetradecyl isocyanate (MW = 239, d = 0.869, 105 µl = 0.38 mmol, i.e., 3 times excess per hydroxyl group present on the solid support), and finally 210 µl of dibutyltin dilaurate (MW = 631.6, d = 1.053, 210 µl = 0.35 mmol). The reaction mixture was purged with nitrogen for approximately 5 min at room temperature and mixed overnight (Intelli-mixer, RM-2, using program F26). The reaction mixture was transferred to a 50 mL tube and chloroform was added to bring the volume to 50 mL. After sonication for approximately 5 min, the white precipitate formed during the reaction dissolved. The solid support was washed with DMF and acetonitrile, and the final product obtained after cleavage of the support was purified by HPLC.

[0407] Scheme 2. Synthesis of compounds A115 and A116 Synthesis of compounds A117 and A118.

[0408] Compounds A117 (X=S(=O)) and A118 (X=S(=O)2) were prepared via a synthetic route similar to that described above for compound A115, with the ethyl methyl sulfide scavenger omitted in the urethane formation step, and optionally nitrogen bubbling omitted. The omission of the ethyl methyl sulfide scavenger yielded a mixture of compounds A115, A117, and A118, which were then separated and purified by HPLC.

[0409] Alternatively, sulfone or sulfoxide derivatives (e.g., A117 and A118) can be prepared by oxidizing the corresponding sulfides (e.g., A115) with oxidants such as m-chloroperoxybenzoic acid (MCPBA) or tert-butylhydroperoxide (t-BuOOH) under appropriate conditions.

[0410] Synthesis of compounds A203 and A204.

[0411] The synthesis of compounds A203 and A204 is described in Scheme 3 below.

[0412] Fmoc-Gly was added as the first amino acid to a solid support, and then coupled with a 2-molar excess of Fmoc-NHCH2CH2O-(PEG) in 2 ml of dimethylformamide (DMF) in the presence of 2-fold excess of benzotriazole hexafluorophosphate tetramethylurea (HBTU), hydroxybenzotriazole (HOBT), and 4-fold excess of diisopropylethylamine (DIPEA). 11 -CH2CH2COOH or Fmoc-NHCH2CH2O-(PEG) 27 -CH2CH2COOH was incubated for 2 hours. Then, Fmoc-Ser(tBu)-OH was coupled to provide intermediate A2, followed by coupling with Boc-Cys(StBu)A1. The thiol tert-butyl group on the cysteine ​​residue was removed by incubating the peptide resin in 0.5 M dithiothreitol in DMF at room temperature (for 1 hour). 250 mg of saturated Boc-Cys-Ser(tBu)-NHCH2CH2O-(PEG) in DMF was then added. 11 -CH2CH2C(O)Gly resin or Boc-Cys-Ser(tBu)CH2CH2O-(PEG) 27 250 µl of R-(+)-1,2-epoxy-butane-4-ol [(R)-2-(epoxyethylene-2-yl)ethane-1-ol] (M) was added to -CH2CH2C(O)-Gly resin (0.25 mmol / g, 0.25 g = 0.0625 mmol). W = 88.11, d = 1.1, 250 µl = 3.125 mmol (equivalent to 50 times the excess of free thiol groups present on the peptide resin) and 25 µl of diisopropylethylamine (DIPEA, M W = 129.2, d = 0.74, 25 µl = 0.14 mmol). The reaction mixture was placed in a water bath at 50°C for 2 hours and then thoroughly washed with DMF to provide intermediate A3.

[0413] Palmitic acid (320 mg, 1.25 mmol), DIPCDI (225 μL, 1.5 mmol), and 4-dimethylaminopyridine (DMAP; 15.25 mg, 0.125 mmol) were dissolved in 2 mL of dichloromethane (DCM) at room temperature and then added to resin-bound BOC-Dhc-peptide resin A3 (0.0625 mmol, 0.25 g) and shaken for 16 h. Prior to undergoing the lysis process described below, the supernatant was removed by filtration, and the solid support was thoroughly washed with DCM and dimethylformamide (DMF) to remove any residual urea.

[0414] The solid support containing the assembled lipopeptide was exposed to reagent B (93% TFA, 5% water, and 2% triisopropylsilane) for 2 hours. To separate the product, most of the TFA was removed, and the residue was then dissolved in 50% acetonitrile and purified immediately using the purification protocol described below, or the material was freeze-dried and stored for subsequent purification.

[0415] Scheme 3. Synthesis of compounds A203 (x = 11) and A204 (x = 27). Synthesis of compounds A215 and A216.

[0416] The synthesis of compounds A215 and A216 was carried out as described in Scheme 4. Intermediate A3 was prepared as described for compounds 3 and 4 above.

[0417] Then, 100 µl of ethyl methyl sulfide (M) was added to 250 mg of peptide resin after glycidization and washing with toluene. W = 76.16, d = 0.842, 100 µl = 1.10 mmol), followed by the addition of 105 µl of tetradecyl isocyanate (MW = 239, d = 0.869, 105 µl = 0.38 mmol, i.e., 3 times excess of each hydroxyl group present on the solid support), and finally the addition of 210 µl of dibutyltin dilaurate (M W = 631.6, d = 1.053, 210 µl = 0.35 mmol). The reaction mixture was purged with nitrogen for approximately 5 min at room temperature and mixed overnight (Intelli-Mixer, RM-2, using program F26). The reaction mixture was transferred to a 50 mL tube and chloroform was added to 50 mL. After sonication for approximately 5 min, the white precipitate formed during the reaction dissolved. The solid support was washed with DMF and acetonitrile, and the final product obtained after cleavage of the support (as above) was purified by HPLC.

[0418] Scheme 4. Synthesis of compounds A215 (x = 11) and A216 (x = 27) Synthesis of A220. Compound A220 was synthesized from Fmoc-RINK MBHA PS resin using standard Fmoc solid-phase peptide synthesis. Fmoc groups were removed after each coupling using 20% ​​piperidine in DMF. Couplings of Fmoc-Gly-OH (2-fold excess) and Fmoc-NH-PEG were performed in DMF using equivalence excess ethyl cyano(oxime) acetate (Oxyma Pure) and diisopropylcarbodiimide (DIC) as coupling agents. 28 Coupling of -CH2CH2COOH (1.4 times excess), Fmoc-Ser(tBu)-OH (2 times excess), and N-(Boc)-S-((R)-2,3-dihydroxybutyl)-L-cysteine ​​(1.5 times excess) was performed at room temperature using tetradecyl chloroformate (12 equivalents, relative to resin moles) and DIEA (24 equivalents, relative to resin moles). This coupling was repeated three times (re-coupling). The first coupling was performed at room temperature using tetradecyl chloroformate (12 equivalents, relative to resin moles) and NMM (24 equivalents, relative to resin moles) in anhydrous DCM / THF (85 / 15), for 18 hours. A second coupling was performed at room temperature using tetradecyl chloroformate (6 equivalents, relative to the number of moles of resin) and NMM (12 equivalents, relative to the number of moles of resin) in anhydrous DCM / THF (85 / 15) for 41 hours. Finally, a third coupling was performed at room temperature using tetradecyl chloroformate (6 equivalents, relative to the number of moles of resin) and NMM (12 equivalents, relative to the number of moles of resin) in anhydrous DCM / THF / toluene (85 / 15 / 5) for 21.5 hours.

[0419] Peptide cleavage from resin, removal of the N-terminal Boc group, and deprotection of the serine side chain were achieved by exposing the resin to a solution of 93% trifluoroacetic acid (TFA), 5% H2O, and 3% triisopropylsilane (TIPS) for 1.5 hours. After the cleavage reaction, the mixture was evaporated, and the resulting residue was redissolved in 30% acetonitrile / water and lyophilized.

[0420] Synthesis of A224. Compound A224 was synthesized from a chlorotriphenylmethyl chloride resin with an initial substitution of 1.6 meq / g via standard Fmoc solid-phase peptide synthesis. First, the first amino acid, Fmoc-Gly-OH, was loaded onto the resin using a 0.5-fold molar excess of Fmoc-Gly-OH and DIEA (1.5-fold excess). This was followed by capping with DMF / MeOH / DIEA (80 / 10 / 10) and Fmoc deprotection to obtain a dry-loaded H-Gly-CT resin with a final substitution of 0.67 meq / g. The Fmoc group was removed after each coupling using 20% ​​piperidine in DMF. Fmoc-NH-PEG was synthesized in DMF using (7-azabenzotriazol-1-yloxy)tripyrrolidinephosphonium hexafluorophosphate (PyAOp; 1.4 equivalents) and diisopropylethylamine (DIEA; 3.2 equivalents). 28 Coupling with -CH2CH2COOH (1.4 equivalents) was performed, while in DMF, a considerable excess of Oxyma Pure and DIC were used as coupling agents for Fmoc-Ser(tBu)-OH (2 equivalents) and N-(Boc)-S-((R)-2,4-dihydroxybutyl)-L-cysteine ​​(1.5 equivalents). Palmitic acid coupling was performed at room temperature using palmitic acid (20 equivalents, relative to resin moles), DIC (20 equivalents), and DMAP (2 equivalents) in DCM / THF (85 / 15) (v / v) for 24 hours.

[0421] Peptide cleavage from resin, removal of the N-terminal Boc group, and deprotection of the serine side chain were achieved by exposing the resin to a solution of 93% TFA, 5% H2O, and 3% TIPS for 1.5 hours. After the cleavage reaction, the mixture was evaporated, and the resulting residue was redissolved in 30% acetonitrile / water and lyophilized.

[0422] Purification and Characterization Purification and characterization: After lysis from the solid support, each analogue was purified by reversed-phase HPLC according to either scheme A or scheme B as described below.

[0423] Option A: Reversed-phase HPLC was performed using an Agilent Zorbax 300SB-C3, 5 μm column (9.4 mm × 250 mm; Agilent Technologies, Santa Clara, California, USA) installed in an Agilent HPLC 1260 Infinity system (Agilent Technologies, Santa Clara, California, USA), and chromatograms were generated using buffer A (0.1% trifluoroacetic acid in water) and buffer B (0.1% trifluoroacetic acid in acetonitrile).

[0424] Option B: Reversed-phase chromatography was performed using a Novasep axial compression column (5 cm diameter) loaded with cyano-based media (Daisogel SP-120-CN-P) and an acetonitrile gradient in [0.1% TFA / water]. After lyophilizing the intermediate, it was ion-exchanged on a Dowex ion-exchange resin to obtain the peptide as an acetate.

[0425] The target materials were identified and their purity determined using an inline HPLC-MS system under the following conditions: Condition A: HPLC column: Agilent Zorbax 300-SB C3 (150 × 0.5 mm; 5 μm) with the following gradient conditions: 0–5 min, 20% B; 5–32 min, 20% B – 100% B; 32–40 min, 100% B – 20% B. Flow rate: 20 μl / min. LC-MS: Agilent 1100 series capillary LC system inlined with an Agilent 1100 series LC / MSD ion trap mass spectrometer. The mass spectrometer was operated using electro-jet ionization set to cation mode. Charged ion series were deconvolved using data analysis software from Agilent Technologies for identification of peptide materials, and the materials were then characterized by LC-MS.

[0426] Condition B: Analytical reversed-phase HPLC with a cyano column (Daiso Fine Chem, SP-120-3-CN-P, 150 × 4.6 mm, 3 µm, 120 Å). Peptides were also analyzed by cation-mode ESI LC-MS using a Finnigan LCQ Deca XPMax.

[0427] The compounds A107, A108, A115, A116, A203, A204, A215 and A216, prepared and purified according to scheme A and condition A as described above, as well as compounds A220 and A224, prepared and purified according to scheme B and condition B as described above, were found to have a purity greater than 95%.

[0428] Peptide quantification Quantification of compounds A107, A108, A115, A116, A203, A204, A215, and A216 was performed by vacuum hydrolysis of the samples in sealed glass vials at 110°C in the presence of 6N HCl containing 0.1% phenol. Amino acid derivatization was then performed using Waters AccQTag reagents according to the manufacturer's instructions, followed by analysis on a Waters Acquity UPLC system (Millipore) using an AccQTag ultracolumn (2.1 mm × 100 mm). Quantification of other compounds was achieved using a similar protocol.

[0429] Synthesis of sulfone and sulfoxide analogs of compounds A215 and A216 Sulfone and sulfoxide derivatives of compounds A215 and A216 can be obtained via a synthetic route similar to that described above, except that the ethyl methyl sulfide scavenger is omitted from the carbamate formation step, and optionally, nitrogen purging is omitted. This reaction yields a mixture of thiols, sulfones, and sulfoxide derivatives, which can be separated and purified by HPLC.

[0430] Alternatively, sulfone or sulfoxide derivatives can be prepared by oxidation of the corresponding sulfides with oxidants such as m-chloroperoxybenzoic acid (MCPBA) or tert-butyl hydroperoxide (t-BuOOH) under appropriate conditions.

[0431] Activation of human TLR2 The potency of these compounds as human and mouse TLR-2 activators was tested in an in vitro assay. This assay assessed NF-κB activation in the HEKBlue-mTLR-2 cell line. These cells were stably transfected with mouse TLR-2 and expressed TLR-1 and TLR-6 endogenously at levels sufficient to allow full functional activation of TLR-1 / 2 and TLR-2 / 6.

[0432] Toll-like receptor 2 (TLR2) stimulation was tested by evaluating NF-κB activation in the HEKBlue-hTLR2 cell line. These cells were stably transfected with human TLR2 and endogenously expressed TLR1 and TLR6 at levels sufficient to allow full functional activation of TLR1 / 2 and TLR2 / 6. The activity of the test sample as a potential agonist against human TLR2 was tested. The test sample was evaluated at seven concentrations and compared with control ligands. These steps were performed in triplicate.

[0433] NF-κB reporter gene assay protocolThis assay was performed as previously described (Jackson et al., 2004; Lau et al., 2006; Sandor et al., 2003; Zeng et al., 2010). HEK293T cells were cultured at 4 × 10⁶ cells per well. 4 Cells were cultured at a density of 100 ng of NF-κB luciferase reporter gene [50 ng of TK-Hyne-Luciferase expression plasmid (Promega Corporation, Madison, USA)] with or without 5 ng of TLR2-expression plasmid in the presence of 0.8 µl Fugene 6 (Roche Diagnostic) for 24 hours. Compounds were added to wells at the concentrations indicated in the histogram 24 hours later. Cell lysates were prepared 5 hours after stimulation using reporter gene lysis buffer (Promega Corporation, Madison, USA). Cell lysates were collected using a kit (Promega Corporation, Madison, USA) and a FLUOstar microplate reader (BMG Labtech, Ottenberg, Germany). Labtech measured luciferase activity in cell lysates. NF-κB-dependent firefly luciferase activity was normalized using NF-κB-independent Renal luciferase activity. Relative stimulation was calculated as the ratio of stimulated to unstimulated samples.

[0434] The results of this determination for compounds A107, A108, A115, A116, A203, A204, A215, and A216 are shown in... Figure 9 These data show that these compounds exhibit significant activity against TLR2.

Claims

1. A method of treating, preventing or minimising progression of cancer in a subject, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby treating, preventing or minimising progression of cancer in the subject.

2. A method of treating, preventing or minimising progression of cancer in a subject who has received or is receiving an immunostimulant, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist, thereby treating, preventing or minimising progression of cancer in the subject.

3. A method of treating, preventing or minimising progression of cancer in a subject, the method comprising the steps of: - identifying a subject with cancer who has received or is receiving an immunostimulant for the treatment of cancer, - assessing whether the subject is responsive to the immunostimulant, - administering to the subject a therapeutically effective amount of a TLR2 agonist if the subject is not responsive to the immunostimulant, thereby treating, preventing or minimising progression of cancer in the subject.

4. A method of treating, preventing or minimising progression of cancer in a subject, the method comprising the steps of: - identifying a subject with cancer who is not responsive to a treatment comprising an immunostimulant, - administering to the subject a therapeutically effective amount of a TLR2 agonist, thereby treating, preventing or minimising progression of cancer in the subject.

5. A method of treating, preventing or minimising progression of cancer in a subject, the method comprising the steps of: - identifying a subject with cancer; and - administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby treating, preventing or minimising progression of cancer in the subject.

6. A method of increasing survival of a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby increasing survival of the subject with cancer.

7. A method of minimising, reducing or preventing tumour growth in a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby minimising, reducing or preventing tumour growth in the subject with cancer.

8. A method of minimising, reducing or preventing metastasis in a subject with cancer, the method comprising administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby minimising, reducing or preventing metastasis in the subject with cancer.

9. The method of claim 8, wherein the method minimises, reduces or prevents metastasis to the lung.

10. A method of minimising, reducing or preventing cancer in a subject, the method comprising: - identifying a subject with a tumour capable of metastasis; and - administering to the subject a therapeutically effective amount of a TLR2 agonist and an immunostimulant, thereby minimising, reducing or preventing cancer in the subject.

11. Use of a TLR2 agonist and an immunostimulant in the manufacture of a medicament for treating, preventing or minimising progression of cancer in a subject. ​ ​ 12. Use of a TLR2 agonist and an immunostimulant for treating, preventing or maximising reduction in progression of cancer in a subject.

13. The method or use of any one of claims 1 to 12, wherein the immunostimulant is selected from the group consisting of cellular immunotherapy, oncolytic virus, cancer vaccine, T cell engaging agent, bispecific T cell engaging agent and checkpoint inhibitor.

14. The method or use of claim 13, wherein the immunostimulant is a checkpoint inhibitor.

15. The method or use of claim 14, wherein the checkpoint inhibitor is a PD-1, PD-L1 or CTLA-4 checkpoint inhibitor, preferably in the form of an antibody.

16. The method or use of claim 15, wherein the checkpoint inhibitor is a PD-1 antibody.

17. The method or use of any one of claims 1 to 16, wherein the cancer is selected from the group consisting of breast cancer, colorectal cancer, adenocarcinoma, mesothelioma, bladder cancer, prostate cancer, germ cell cancer, hepatocellular carcinoma / cholangiocellular carcinoma, neuroendocrine cancer, pituitary tumour, small round cell tumour, squamous cell carcinoma, melanoma, atypical fibroxanthoma, seminoma, non-seminoma, interstitial Leydig cell tumour, Sertoli cell tumour, skin tumour, kidney tumour, testicular tumour, brain tumour, ovarian tumour, gastric tumour, oral cavity tumour, bladder tumour, bone tumour, fibrosarcoma, cervical tumour, oesophageal tumour, laryngeal tumour, liver tumour, lung tumour, vaginal tumour or Wilms' tumour; in preferred embodiments, the cancer is melanoma or colon cancer.

18. The method or use of claim 17, wherein the cancer is melanoma, breast cancer, fibrosarcoma or colon cancer.

19. The method or use of any one of claims 1 to 18, wherein the TLR2 agonist is administered in a composition further comprising a pharmaceutically acceptable carrier, diluent or excipient.

20. The method or use of claim 19, wherein the composition consists of a TLR2 agonist, an immunostimulant and a pharmaceutically acceptable carrier, diluent or excipient.

21. The method or use of any one of claims 1 to 20, wherein the TLR2 agonist comprises a lipid, a peptidoglycan, a lipoprotein or a lipopolysaccharide.

22. The method or use of any one of claims 1 to 21, wherein the TLR2 agonist comprises Pam2Cys-Ser-PEG.

23. The method or use of any one of claims 1 to 22, wherein the checkpoint inhibitor is a PD-1 antibody and the TLR2 agonist comprises Pam2Cys-Ser-PEG.

24. The method or use of any one of claims 1 to 23, wherein the TLR2 agonist comprises palmitoyl, myristoyl, stearoyl, lauroyl, octanoyl or decanoyl.

25. The method or use of any one of claims 1 to 21, wherein the TLR2 agonist is selected from the group consisting of Pam2Cys, Pam3Cys, Ste2Cys, Lau2Cys, and Oct2Cys.

26. The method or use of any one of claims 1 to 21, wherein the TLR2 agonist comprises Pam2Cys.

27. The method or use of any one of claims 1 to 26, wherein the compound comprises a TLR2 agonist and a solubilizer.

28. The method or use of claim 27, wherein the TLR2 agonist is linked to the solubilizer.

29. The method or use of claim 27 or 28, wherein the solubilizer comprises or consists of a positively or negatively charged group.

30. The method or use of claim 29, wherein the charged group is a branched or linear peptide.

31. The method or use of claim 29, wherein the positively charged group comprises at least one positively charged amino acid, preferably an arginine or lysine residue.

32. The method or use of claim 29, wherein the negatively charged group comprises at least one negatively charged amino acid, preferably a glutamic acid or aspartic acid.

33. The method or use of claim 30, wherein the branched or linear peptide is R4, H4, H8, or E8.

34. The method or use of claim 30, wherein the branched peptide comprises 。 35. The method or use of claim 27, wherein the solubilizer comprises polyethylene glycol (PEG) or R4.

36. The method or use of claim 27, wherein the solubilizer comprises polyethylene glycol (PEG) and R4.

37. The method or use of claim 35 or 36, wherein the PEG is PEG 11 or PEG 12 .

38. The method or use of any one of claims 1 to 34, wherein the TLR2 agonist is a compound of formula (I): ， wherein A comprises or consists of a moiety selected from A1 and A2: ， wherein each z is independently selected from 1 or 2; each X is independently selected from -S-, -S(=0)-, and -S(=0)2-; in moiety A1: each g is independently 10, 11, 12, 13, 14, 15, 16, 17, or 18; R6 and R7 are independently selected from the group consisting of H, linear or branched C1-C4 alkyl, and -C(=0)CH3; R9and R 10 are independently selected from the group consisting of -NH-, -O-, or a single bond; and in moiety A2: b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, provided that: the sum of b, v, and w is at least 3; and the sum of b and w is from 0 to 7; Z1 and Z2 are each independently selected from the group consisting of -0-, -NR-, -S-, S(=0), -S(=0)2-, -C(=0)0-, -OC(=0)-, -C(=0)NR-, -NRC(=0)-, -C(=0)S-, -SC(=0)-, -OC(=0)0-, -NRC(=0)0-, -OC(=0)NR-, and -NRC(=0)NR-; R 11 , R 12 , R x , R y , R 14 , R 15 , R 16 and R 17 are each independently H or C1-C6 aliphatic; R, R 13 and R 18 each independently is H or C1-C6aliphatic; R 19 is H, C1-C6aliphatic, an amino protecting group, L3-C(=0)-, or A2; L1and L2are each independently C5-C 21 aliphatic or C4-C 20 heteroaliphatic; L3is C1-C 21 aliphatic or C2-C 20 heteroaliphatic; A2 is an amino acid or a peptide; wherein any aliphatic or heteroaliphatic group present in R, R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R x , R y , L1, L2, and L3 is optionally substituted; Y is ， R1and R2are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=0)NH2, -CH2CH2C(=0)OH, and -CH2CH2C(=0)OR8, wherein any of the alkyl hydrogens can be replaced with a halogen; R8is selected from the group consisting of H and linear or branched C1-C6alkyl; and B comprises or consists of polyethylene glycol (PEG), or a pharmaceutically acceptable salt or prodrug thereof.

39. The method or use of any one of claims 1 to 34, wherein the compound comprises or consists of the partial structure A1Y’ or A2Y’: (A1Y') (A2Y') R1and R2are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=0)NH2, -CH2CH2C(=0)OH, and -CH2CH2C(=0)OR8, wherein any of the alkyl hydrogens can be replaced with a halogen; R6and R7are independently selected from the group consisting of H, linear or branched C1-C4alkyl, and -C(=0)CH3; R8is selected from the group consisting of H and linear or branched C1-C6alkyl; R9and R 10 are independently selected from the group consisting of -NH-, -0-, or a single bond; z is 1 or 2; X is selected from -S-, -S(=0)-, and -S(=0)2-; b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, provided that: the sum of b, v, and w is at least 3; and the sum of b and w is from 0 to 7; Z1and Z2are each independently selected from the group consisting of -0-, -NR-, -S-, -S(=0)-, -S(=0)2-, -C(=0)0-, -OC(=0)-, -C(=0)NR-, -NRC(=0)-, -C(=0)S-, -SC(=0)-, -OC(=0)0-, -NRC(=0)0-, -OC(=0)NR-, and -NRC(=0)NR-; In each instance of b, v, w, and z, R 11 , R 12 , R x , R y , R 14 , R 15 , R 16 , and R 17 are each independently H or C1-C6 aliphatic; R, R 13 and R 18 each independently is H or C1-C6 aliphatic; R 19 is H, C1-C6aliphatic, an amino protecting group, L3-C(=0)-, or A2; L1and L2are each independently C5-C 21 aliphatic or C4-C 20 heteroaliphatic; L3is C1-C 21 aliphatic or C2-C 20 heteroaliphatic; A2is an amino acid or a peptide; wherein any aliphatic or heteroaliphatic group present in R, R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R x , R y , L1, L2, and L3 is optionally substituted; and A1Y’ or A2Y’ is covalently linked to polyethylene glycol (PEG), or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

40. The method or use of any one of claims 1 to 34, wherein the TLR2 agonist is a compound comprising a moiety A selected from A1’ and A2and PEG, the moiety A and PEG are linked through a glycine, serine, homoserine, threonine, phosphoserine, asparagine, or glutamine residue or an ester of a glutamine residue ， wherein: each g is independently 10, 11, 12, 13, 14, 15, 16, 17, or 18 each z is independently selected from 1 or 2; each X is independently selected from -S-, -S(=0)-, and -S(=0)2-; b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, provided that: the sum of b, v, and w is at least 3; and the sum of b and w is from 0 to 7; Z1and Z2are each independently selected from the group consisting of -0-, -NR-, -S-, S(=0), -S(=0)2-, -C(=0)0-, -OC(=0)-, -C(=0)NR-, -NRC(=0)-, -C(=0)S-, -SC(=0)-, -OC(=0)0-, -NRC(=0)0-, -OC(=0)NR-, and -NRC(=0)NR-; R 11 , R 12 , R x , R y , R 14 , R 15 , R 16 and R 17 are each independently H or C1-C6 aliphatic; R, R 13 and R 18 each independently is H or C1-C6aliphatic; R 19 is H, C1-C6aliphatic, an amino protecting group, L3-C(=0)-, or A2; L1and L2are each independently C5-C 21 aliphatic or C4-C 20 heteroaliphatic; L3is C1-C 21 aliphatic or C2-C 20 heteroaliphatic; A2is an amino acid or a peptide; wherein any aliphatic or heteroaliphatic group present in R, R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R x , R y , L1, L2, and L3 is optionally substituted or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

41. The method or use of any one of claims 1 to 34, wherein the TLR2 agonist is a compound of Formula (VIII): ， wherein A is a moiety selected from A1and A2 ， wherein each z is independently selected from 1 or 2; each X is independently selected from -S-, -S(=0)-, and -S(=0)2-; In moiety A1: each g is independently 10, 11, 12, 13, 14, 15, 16, 17, or 18; R6and R7are independently selected from the group consisting of H, straight or branched C1-C4alkyl, and -C(=0)CH3; R9and R 10 are independently selected from the group consisting of -NH-, -O-, or a single bond; and In moiety A2: b and w are each independently an integer from 0 to 7, and v is an integer from 0 to 5, such as from 2 to 5, provided that: the sum of b, v, and w is at least 3; and the sum of b and w is from 0 to 7; Z1and Z2are each independently selected from the group consisting of -0-, -NR-, -S-, S(=0), -S(=0)2-, -C(=0)0-, -OC(=0)-, -C(=0)NR-, -NRC(=0)-, -C(=0)S-, -SC(=0)-, -OC(=0)0-, -NRC(=0)0-, -OC(=0)NR-, and -NRC(=0)NR-; R 11 , R 12 , R x , R y , R 14 , R 15 , R 16 and R 17 are each independently H or C1-C6 aliphatic; R, R 13 and R 18 each independently is H or C1-C6aliphatic; R 19 is H, C1-C6aliphatic, an amino protecting group, L3-C(=0)-, or A2; L1and L2are each independently C5-C 21 aliphatic or C4-C 20 heteroaliphatic; L3is C1-C 21 aliphatic or C2-C 20 heteroaliphatic; A2is an amino acid or a peptide; wherein any aliphatic or heteroaliphatic group present in R, R 11 , R 12 , R 13 , R 14 , R 15 , R 16 , R 17 , R 18 , R 19 , R x , R y , L1, L2, and L3 is optionally substituted; Y is ， wherein R1and R2are independently selected from the group consisting of H, -CH2OH, -CH2CH2OH, -CH(CH3)OH, -CH2OPO(OH)2, -CH2C(=0)NH2, -CH2CH2C(=0)OH, and -CH2CH2C(=0)OR8, wherein any of the alkyl hydrogens can be replaced with a halogen; R6and R7are independently selected from the group consisting of H, straight or branched C1-C4alkyl, and -C(=0)CH3; R8is selected from the group consisting of H and straight or branched C1-C6alkyl; R9and R 10 are independently selected from the group consisting of -NH-, -O-, or a single bond; n is 3 to 100; m is 1, 2, 3, or 4; p is 2, 3, or 4; q is zero or 1; wherein when q = 1, R3is -NH2or -OH; wherein when q = 0, R3is H; L is null or consists of 1 to 10 units, wherein each unit is a natural alpha amino acid or is derived from a natural alpha amino acid, and has the following formula: ， wherein R4is H; and R5is a side chain, or a second hydrogen of the amino acid, or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

42. The method or use of any one of claims 1 to 34, wherein the TLR2 agonist is a compound selected from the following table: or a pharmaceutically acceptable salt, solvate, or prodrug thereof.

43. The method or use of claim 43, wherein the TLR2 agonist is defined by the formula: ， and the immune stimulator is a PD-1, PD-L1, or CTLA-4 antibody.

44. The method or use of claim 43, wherein the TLR2 agonist is defined by the formula: ， and the immune stimulator is a PD-1, PD-L1, or CTLA-4 antibody.

45. The method or use of any one of claims 1 to 45, wherein the TLR2 agonist is not Pam3Cys.

46. The method or use of any one of claims 1 to 46, wherein the therapeutically effective amount of the TLR2 agonist and immune stimulator is administered once per day.

47. The method or use of any one of claims 1 to 46, wherein the therapeutically effective amount of the TLR2 agonist and immune stimulator is administered once per week.

48. The method or use of any one of claims 1 to 48, wherein the therapeutically effective amount of the TLR2 agonist and immune stimulator is adapted for intravenous administration to the subject.

49. The method or use of any one of claims 1 to 48, wherein the therapeutically effective amount of the TLR2 agonist and immune stimulator is adapted for administration to the respiratory tract, preferably by inhalation.

50. A TLR2 agonist for use in combination with an immune stimulator in the treatment, prevention, or maximal reduction of cancer progression in a subject.

51. An immune stimulator for use in combination with a TLR2 agonist in the treatment, prevention, or maximal reduction of cancer progression in a subject.

52. Use of a TLR2 agonist in combination with an immune stimulator in the treatment, prevention, or maximal reduction of cancer progression in a subject.

53. A pharmaceutical composition comprising a therapeutically effective amount of a TLR2 agonist and an immune stimulator for use in the treatment, prevention, or maximal reduction of cancer progression in a subject.

54. The TLR2 agonist of claim 51, the immune stimulator of claim 52, the use of claim 53, or the composition of claim 54, wherein the TLR2 agonist is Pam2Cys-Ser-PEG defined by the formula: ， and the immune stimulator is a PD-1, PD-L1, or CTLA-4 antibody.

55. The TLR2 agonist of claim 51, the immune stimulator of claim 52, the use of claim 53, or the composition of claim 54, wherein the TLR2 agonist is defined by the formula: ， and the immune stimulator is a PD-1, PD-L1, or CTLA-4 antibody.

56. The TLR2 agonist, immune stimulator, use, or pharmaceutical composition of any one of claims 51 to 56, wherein the therapeutically effective amount of the TLR2 agonist and immune stimulator is adapted for administration to the respiratory tract of the subject, preferably by inhalation.

57. The TLR2 agonist, immune stimulator, use, or pharmaceutical composition of claim 57, wherein the composition is formulated as a nasal spray or nasal drops.

58. The method, use, TLR2 agonist, immunostimulant or pharmaceutical composition of any one of claims 1 to 58, wherein the TLR2 agonist increases the effectiveness of the immunostimulant in the subject.

59. The method, use, TLR2 agonist, immunostimulant or pharmaceutical composition of claim 59, wherein the TLR2 agonist increases survival, reduces tumor growth and / or reduces metastasis of the tumor in the subject.

60. A kit comprising a TLR2 agonist of any one of claims 13 to 16 or 19 to 46 and / or a checkpoint inhibitor for use in treating, preventing or minimizing progression of cancer in a subject.

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