CPG Amphifil and its Use

Abstract

The present invention provides compounds comprising a CpG oligodeoxynucleotide sequence linked to a lipid by a linker, as well as related compositions and methods. [Selected Figure] Figure 1

Description

Array list

[0001] This application includes a sequence listing submitted electronically in ASCII format, which is hereby incorporated by reference in its entirety. The name of the ASCII copy created by _ is _, and its size is _ bytes.

Background Art

[0002] Human papillomavirus (HPV)-associated cancers are one of the cancers with the most rapid increase in the world. Overall, it is considered that 5% of all cancers in the world are caused by HPV infection. Further effective cancer treatments are required.

Summary of the Invention

[0003] The present invention provides a compound that can be used in a treatment method.

[0004] That is, in a first aspect, the present invention provides the following lipid

Chemical formula

[0005] In one embodiment of the first aspect of the present invention, the nucleotide sequence is bound to the lipid. <00所<3000028> In another embodiment of the first aspect of the present invention, all internucleoside groups linking the nucleosides of 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3' (SEQ ID NO: 1) are phosphorothioates.

[0007] In a second aspect, the present invention is characterized by a method for treating cancer in human patients. This method comprises administering to a patient the compound of the first aspect, a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

[0008] In one embodiment of a second aspect of the present invention, the cancer is positive for human papillomavirus (HPV) (for example, positive for HPV type 16).

[0009] In another embodiment of a second aspect of the present invention, the cancer is head and neck squamous cell carcinoma.

[0010] In an additional embodiment of the second aspect of the present invention, the patient is or has been receiving platinum-containing chemotherapy. In a further embodiment, an anti-PD-1 antibody (e.g., pembrolizumab or nivolumab) is administered to the patient.

[0011] In another embodiment of the second aspect of the present invention, the compound of the first aspect of the present invention, and the protein having the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO: 3 are administered simultaneously.

[0012] In a further embodiment of the second aspect of the present invention, the compound of the first aspect of the present invention, and the protein having the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO: 3 are administered sequentially.

[0013] In a further aspect, the present invention features an alternative method for treating cancer in human patients. This method comprises administering to a patient a compound of the first aspect of the present invention, a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3), and an anti-PD-1 antibody (e.g., pembrolizumab or nivolumab).

[0014] In a third aspect, the present invention is characterized by a pharmaceutical composition comprising the compound of the first aspect of the present invention and a pharmaceutically acceptable carrier.

[0015] In one embodiment of the third aspect, the pharmaceutical composition further comprises a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

[0016] In a fourth aspect, the present invention is characterized by (i) a compound of the first aspect of the present invention, or a composition of the second aspect of the present invention, and (ii) a kit comprising a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

[0017] definition As used herein, "linker" refers to a monovalent or divalent group that is covalently bonded to one biological functional group with one valency and to another biological functional group with the other valency. For example, a linker can link the nucleotide sequence of a CpG oligonucleotide to a lipid (as described herein, for example, -P(X)(OH)-O-CH(CH2NHCO-(CH2) 16 The linker is a -CH3)2 or a salt thereof, where X is O or S. Such a linker may optionally contain one or more nucleotides, for example, a dinucleotide (e.g., GG).

[0018] As used herein, “pharmaceutically acceptable carrier” refers to a vehicle capable of suspending or dissolving an active compound, which is non-toxic and non-inflammatory to the patient. Furthermore, a pharmaceutically acceptable carrier may include pharmaceutically acceptable additives such as preservatives, antioxidants, fragrances, emulsifiers, dyes, or excipients, which are known or used in the field of pharmaceutical formulations, and which do not significantly inhibit the therapeutic effect of the biological activity of the active agent and are non-toxic to the patient.

[0019] The terms “treat,” “treatment,” and “treating” refer to therapeutic approaches aimed at reversing, mitigating, improving, inhibiting, slowing, or halting the progression or severity of a condition associated with a disease or disorder, such as cancer. These terms include reducing or alleviating at least one adverse effect or symptom of a condition, disease, or disorder. A treatment is generally “effective” if one or more symptoms or clinical markers are reduced or a desired response (e.g., a specific immune response) is induced. Alternatively, a treatment is “effective” if the progression of the disease is reduced or halted.

[0020] The present invention offers several advantages. For example, certain compounds of the present invention, which include a lipid moiety and optionally a linker, bind to endogenous albumin in the target to which they are administered, thereby facilitating the delivery of the compounds to the target's lymph nodes. This, for example, facilitates the induction of a therapeutic immune response against the administered HPV protein in the target, leading to effective cancer treatment.

[0021] Other features and advantages of the present invention will become apparent from the following detailed description, drawings, and claims. [Brief explanation of the drawing]

[0022] [Figure 1] Figure 1 is a graph showing the immune response to HPV16 E6. [Figure 2] Figure 2 is a graph showing the immune response to HPV16 E7. [Figure 3] Figure 3 is a graph showing the tetramer staining analysis for HPV16 E7. [Figure 4] Figure 4 is a series of graphs showing that administration of amphiphile-CpG (aCpG) with HPV16 E6 and HPV16 E7 reduced tumor size compared to aCpG alone or no treatment (No Tx). [Figure 5]Figure 5 is a graph showing the excellent HPV tetramer response by the aCpG vaccine compared to the soluble CpG vaccine at different time points. [Figure 6] Figure 6 is a graph showing the persistence of the HPV tetramer response over time when aCpG is administered weekly or once every two weeks. [Figure 7] Figure 7 is a graph showing the response of tumor size to E7 vaccine treatment. [Figure 8] Figure 8 is a graph showing improved survival in C57BL6 mice transplanted with TC-1 tumor cells treated with the E7 vaccine. [Figure 9] Figure 9 is a series of graphs showing changes in serum cytokine levels after administration. [Figure 10] Figure 10 is a graph showing the tetramer response of the HPV16 E7 / aCpG vaccine over time. [Figure 11] Figure 11 is a graph showing the tumor growth response when the administration of an anti-PD-1 antibody is plus or minus in the HPV16 E7 / aCpG vaccine inoculation. [Figure 12] Figure 12 is a graph showing the effect on the survival rate of TC-1 tumor-bearing mice when an anti-PD-1 antibody is plus or minus administered in the HPV16 E7 / aCpG vaccine inoculation. [Figure 13] Figure 13 is a graph showing the tetramer analysis of aCpG dose escalation. [Figure 14] Figure 14 is a graph showing the structure of Amphiphil-CpG-7909; 5'-(diacyl lipid) TCG TTT TGT CGT TTT GTC GTT-3' (SEQ ID NO: 1). All bases are DNA. All linkages, including the linkage between the diacyl lipid and the oligodeoxynucleotide, are phosphoramidites.

Mode for Carrying Out the Invention

[0023] The present invention provides compounds that can be used in therapeutic methods. The compounds include CpG oligodeoxynucleotides (ODNs) (for example, CpG ODNs have the sequence 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3' (SEQ ID NO: 1)). At the 5' end of the CpG ODN, for example, the following [ka] It is linked to a lipid or a salt thereof, such as (where X is O or S). The CpG oligodeoxynucleotide may be directly bound to the lipid, or the CpG may be linked to the lipid via a linker such as GG. In the CpG oligodeoxynucleotide, all internucleoside groups are phosphorothioates (for example, all internucleoside groups of a compound may be phosphorothioates).

[0024] CpG ODNs can function as adjuvants to induce an immune response in a target, such as an immune response to a cancer antigen (e.g., HPV antigen). Thus, the compounds and compositions of the present invention can be used in therapeutic methods. In particular, when a CpG ODN-containing compound is administered in combination with one or more HPV proteins, the compound can induce an immune response against HPV-positive cancer cells. Therefore, the present invention provides a method for treating a cancer in a target (e.g., a human patient) by administering one or more compounds or compositions of the present invention to the target. In various examples, the cancer is HPV-positive (e.g., HPV type 16 positive) cancer.

[0025] HPV-positive cancers include head and neck squamous cell carcinoma, cervical cancer, anal cancer, vulvar cancer, head and neck cancer, oropharyngeal cancer, penile cancer, vaginal cancer, virally induced cancer, bladder cancer, pancreatic cancer, lung cancer, liver cancer, ovarian cancer, colon cancer, stomach cancer, neuroblastoma, breast cancer, prostate cancer, kidney cancer, leukemia, sarcoma, calcinoma, basal cell calcinoma, non-small cell lung calcinoma, non-Hodgkin lymphoma, acute myeloid leukemia (AML), chronic lymphocytic leukemia (CLL), and B-cell chronic lymphocytic leukemia. This may include hematological diseases (B-CLL), multiple myeloma (MM), erythroleukemia, renal cell carcinoma, sarcoma, melanoma, astrocytoma, oligodendrocyte, biliary tract cancer, choriocarcinoma, CNS cancer, laryngeal cancer, small cell lung cancer, non-small cell lung cancer (NSCLC), adenocarcinoma, giant (or auto) cell carcinoma, squamous cell carcinoma, oral cancer, skin cancer, basal cell carcinoma, squamous cell carcinoma, testicular cancer, thyroid cancer, uterine cancer, rectal cancer, respiratory cancer, or urinary tract cancer.

[0026] Optionally, the method of the present invention may further include the step of administering the compound or composition of the present invention in combination with a second (or further) different approach for treatment.

[0027] Furthermore, the present invention provides a kit in which each comprises, for example, a first container containing one or more compounds of the present invention, and optionally a second container containing a cancer antigen such as an HPV protein described herein.

[0028] CpG CpG ODNs are short synthetic single-stranded DNA molecules containing unmethylated CpG dinucleotides in a specific sequence context. CpG ODNs possess a partially or fully phosphorothiolated (PS) backbone, in contrast to the native phosphodiester (PO) backbone in DNA molecules. Based on their structural characteristics and activity in human peripheral blood mononuclear cells (PBMCs), particularly B cells and plasmacytoid dendritic cells (pDCs), three major classes of irritant CpG ODNs have been identified. These three classes are class A (type D), class B (type K), and class C.

[0029] Both CpG1826 and CpG7909 are CpG class B. Class B CpG ODNs contain a full PS backbone with one or more CpG dinucleotides. They strongly activate B cell and TLR9-dependent NF-κB signaling but weakly stimulate IFN-α secretion.

[0030] Mutant HPV The C70G, C113G, and I135G (underlined below) point mutations can be introduced into the wild-type HPV16 E6 viral protein to prevent stereochemical interaction with human p53. The performance of this component as an antigen is determined by the protein sequence, and the protein structure is not important for its intended function. mHPV 16 E6 (158 aa; Sequence ID 2) MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAV G DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC IN G QKPLCPE EKQRHLDKKQ RFHN G RGRWT GRCMSCCRSS RTRRETQL

[0031] The C24G and E26G (underlined below) point mutations can be introduced into the wild-type HPV16 E7 viral protein to prevent stereochemical interaction with human Rb1. Here again, the performance of this component as an antigen is determined by the protein sequence, and the protein structure is not important for its intended function. mHPV 16 E7 (98 aa; Sequence ID 3) MHGDTPTLHE YMLDLQPETT DLY G Y G QLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP.

[0032] CpG ODNs may be directly bound to lipids or linked by linkers. These compounds may be prepared using conventional phosphoramidite chemistry known in the art. In some examples, CpG ODN or CpG ODN-GG is used in the following compounds [ka] The intermediate may be reacted with the intermediate, and the compound of the present invention may be produced by oxidation (for example, a phosphite oxidation method known in the art, for example, a sulfidating agent, for example, 3-((N,N-dimethylaminomethylidene)amino)-3H-1,2,4-dithiazole-5-thion) and hydrolysis of the cyanoethyl group.

[0033] Examples are provided below to better illustrate the present invention. These examples are for illustrative purposes only and should not be construed as limiting the scope of the invention in any way. [Examples]

[0034] Example 1: Generation of an immune response by combining HPV16 E6 and HPV16 E7 proteins with CpG amphifil adjuvant. Mice were prophylactically immunized, and the resulting immune response was recorded via E7-tetramer staining and IFNγ-intracellular cytokine staining (ICS) upon stimulation with HPV16 E6 and HPV16 E7 (E6 / E7).

[0035] The experimental design included the following six groups of mice (n=10 in each group). 1. Non-immunization 2. E6 / E7 + Soluble CpG1826 3. E6 / E7 + Amphibole CpG1826 (aCpG1826) 4. E6 / E7 + Soluble CpG7909 5. E6 / E7 + Amphifil CpG7909 (aCpG7909; Figure 14). 6. E6 / E7 + Poly IC (pIC)

[0036] pIC was used as the benchmark adjuvant control.

[0037] The protein stock solution was dissolved in 8M urea. The adjuvant stock solution was dissolved in H2O. The final injector was 1X phosphate-buffered saline (PBS) (urea C F It is diluted with <1M.

[0038] For aCpG1826, the soluble CpG1826 sequence (5'-tccatgacgttcctgacgtt-3'; SEQ ID NO: 4) with two guanines added to the 5' end was used, as was the sequence (5'-gg tccatgacgttcctgacgtt-3'; SEQ ID NO: 5). Both soluble CpG1826 and aCpG1826 were used at a concentration of 5 nmol in 100 μl injections. CpG1826 is the optimal sequence for mice, while CpG7909 is optimal for humans and exhibits low activity in mice. CpG1826 and CpG7909 belong to the same CpG class (Class B) and generally have similar activity profiles in their respective species.

[0039] For both aCpG7909 and soluble CpG7909, the sequence 5'-tcgtcgttttgtcgttttgtcgtt-3' (SEQ ID NO: 6) was used at a concentration of 5 nmol in each 100 μl injection.

[0040] We used mutant HPV16 E6 with point mutations at C70G, C113G, and I135G (underlined below) for immunization. The amino acid sequence used is shown below. mHPV 16 E6 (158 aa; Sequence ID 2) MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAV G DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC IN GQKPLCPE EKQRHLDKKQ RFHN G RGRWT GRCMSCCRSS RTRRETQL

[0041] We used mutant HPV16 E7 with point mutations at C24G and E26G (underlined below) for immunization. The amino acid sequence used is shown below. mHPV 16 E7 (98 aa; Sequence ID 3) MHGDTPTLHE YMLDLQPETT DLY G Y G QLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP

[0042] For E6 / E7, 10 μg each of mutant HPV16 E6 and mutant HPV16 E7 was used per 100 μl injection.

[0043] Female C57BL / 6J mice (B6) were subcutaneously (sc) immunized with a primer dose (E6 / 7 + aCpG) and a single booster dose two weeks later.

[0044] Tetramer analysis for H-2Db HPV16 E7 (RAHYNIVTF; SEQ ID NO: 7) was performed 7 days after booster administration (Figure 3).

[0045] To analyze the immune response to E6 / E7, intracellular cytokine staining (ICS) for IFNγ was performed in peripheral blood 7 days after booster administration.

[0046] The following peptides were used for E6 stimulation, and the data shown in Figure 1 was generated: E6-10: EVYDFAFRDL (SEQ ID NO: 8), E6 49-57: VYDFAFRDL (SEQ ID NO: 9), E6 37-45: CVYCKQQLL (SEQ ID NO: 10), E6 72-80: KCLKFYSKI (SEQ ID NO: 11), and E6 100-108: NKPLCDLLI (SEQ ID NO: 12).

[0047] Deconvolution of E6 stimulation revealed that E49-57 was the only peptide that provided the stimulation.

[0048] The following peptide was used for E7 stimulation: RAHYNIVTF (SEQ ID NO: 13). Using this peptide, the data shown in Figure 2 was generated.

[0049] As shown in Figures 1 and 2, the use of aCpG1826 induced a strong immune response against both mutant HPV16 E6 and mutant HPV16 E7.

[0050] Furthermore, as shown in Figures 1 and 2, the use of aCpG7909, which is optimal in humans and generally ineffective in mice, surprisingly induced a strong immune response against both mutant HPV16 E6 and mutant HPV16 E7.

[0051] E6 / E7 + aCpG reduced tumor growth and showed a corresponding increase in percentage survival compared to aCpG alone or no treatment (Figure 4).

[0052] Example 2: Determination of the administration schedule for HPV16 E7 and aCPG To determine the optimal dosing schedule for E7+ aCpG regarding its antitumor effect in female C57BL / 6U(B6) mice transplanted with TC-1 tumors, weekly dosing was compared to bi-weekly dosing and baseline (prime only) dosing. E7+ aCpG was compared to E7+ soluble CpG. The entire vaccine was administered in three doses (prime and two boost doses).

[0053] On days 0 and 12, 50,000 TC-1 cells were subcutaneously inoculated into the flanks of female C57BL / 6J mice (B6). The mice were divided into treatment groups and treated as shown in Table 1.

[0054] [Table 1]

[0055] HPV tetramer-specific T-cell responses to protein / amphiphilic CpG vaccines were superior to those of protein / soluble CpG vaccines, both after single and repeated administrations (Figure 5). HPV tetramer responses to protein / aCpG further increased after boost inoculation, and this increase persisted until days 28 and 35 for weekly and bi-weekly regimens, respectively (Figure 6). The strong HPV tetramer response in the aCpG group correlated with reduced tumor size (Figure 7) and improved survival compared to animals vaccinated with soluble CpG (Figure 8). Treatment-related increases in systemic cytokines were similar in the soluble and aCpG groups, except for IL-10 and IFNγ. IL-10 levels were lower in aCpG compared to soluble CpG, and IFNγ levels were higher in aCpG compared to soluble CpG (Figure 9).

[0056] Example 3: Antitumor effect of E7 protein in combination with soluble or amphilic CpG with and without anti-PD-1 antibody. To evaluate the antitumor effect of E7 protein with and without anti-PD-1 antibody, or in combination with amphilic CpG, 50,000 TC-1 cells were inoculated into the flanks of female C57BL / 6J mice (B6) at baseline. Eleven days after inoculation, the mice were divided into five groups as shown in Table 2. The control group received no treatment.

[0057] [Table 2]

[0058] During the study period, tumor size was measured every other day until 40 days after vaccination, and animal survival rates were monitored. Tetramer analysis of H-2Db HPV16 E7(RAHYNIVTF) was performed 7 days after each vaccine administration.

[0059] Administration of E7 / Amph-CpG vaccine with and without anti-PD-1 antibody induced a stable increase in HPV tetramer-+CD8 cells specific to the HPV16 E7(RAHYNIVTF; SEQ ID NO: 13) peptide (Figure 10). This response was clearly observed early after the first dose, peaked with the second dose, and persisted until the third dose (in contrast, the response was low with E7 / CpG, and although it increased with co-administration of anti-PD-1, it was not sustained).

[0060] In response to this robust HPV tetramer + CD8 response, tumor growth stopped around day 24 and reversed after the first dose of E7 / Amph-CpG (with and without anti-PD-1 antibody). In contrast to the other groups where growth progressed, the tumors remained small and stable until the end of the study (Figure 11).

[0061] Furthermore, in relation to the effect on tumor size, treatment with the E7 / aCpG vaccine (with and without anti-PD-1 antibody) had a significant effect on survival rates, with 6 / 7 (85%) cured with E7 / aCpG without antibody and 8 / 10 (80%) cured with E7 / aCpG + anti-PD-1 antibody (Figure 12).

[0062] Example 4: aCpG dose escalation study To determine the dose of aCpG that elicited the maximum antigen-specific tetramer + CD8 response over a course of six administrations, a dose-escalation study was conducted using a fixed dose of 10 μg of ovalbumin (OVA) as the antigen. Soluble CpG was used as a control. Tolerance (based on body weight and general observation) was also evaluated. The study design is outlined in Table 3.

[0063] [Table 3]

[0064] The total trial period was 11 weeks, during which six doses of the vaccine were administered at two-week intervals.

[0065] Peripheral blood samples were collected 7 days after each injection, and flow cytometry analysis of tetramers on CD8+ cells was performed using H-2Kb OVA (SIINFEKL; SEQ ID NO: 14).

[0066] A significant increase in tetramer+CD8+ cells was observed only in the group treated with aCpG+OVA, with 6 nmol yielding the greatest pharmacological effect (Figure 13). No weight loss, decreased interest / appetite, or wounds / injuries were observed.

[0067] Other Embodiments While the present invention has been described in relation to its particular embodiments, further modifications are possible, and it can be understood that this application is generally intended to cover any variations, uses, or adaptations of the present invention in accordance with the principles of the present invention, including any deviations from the disclosure herein that are within the scope of known or customary practice in the art to which the invention belongs and that can be applied to the essential features set forth herein.

[0068] All publications, patents, and patent applications are incorporated herein by whole to the same extent that each individual publication, patent, or patent application is specifically and individually indicated so as to be incorporated by whole.

[0069] Some embodiments of the present invention are described in the following numbered paragraphs.

[0070] 1. The following lipids [ka] Alternatively, a compound comprising a salt thereof and the nucleotide sequence 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3' (SEQ ID NO: 1) linked at the 5' end by a bond or linker (where X is O or S).

[0071] 2. The compound of paragraph 1, wherein the above nucleotide sequence is bound to the above lipid.

[0072] 3. The compound of paragraph 1 or 2, wherein all internucleoside groups linking the nucleoside of 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3' (SEQ ID NO: 1) are phosphorothioates.

[0073] 4. A method for treating cancer in a human patient, comprising the step of administering to the patient one of the compounds described in paragraphs 1 to 3, a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

[0074] 5. The method described in paragraph 4, for the above cancer being human papillomavirus (HPV) positive.

[0075] 6. The above cancer is HPV type 16 positive, using the method described in paragraph 5.

[0076] 7. The above cancer is head and neck squamous cell carcinoma, one of the methods described in paragraphs 4-6.

[0077] 8. The patient described above is currently receiving or has previously received platinum-containing chemotherapy, according to one of the methods described in paragraphs 4-7.

[0078] 9. The method of paragraph 4, wherein the compound of paragraph 1 and the protein having the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO: 3 are administered simultaneously.

[0079] 10. The method of paragraph 4, wherein the compound of paragraph 1 and the protein having the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO: 3 are administered sequentially.

[0080] 11. A pharmaceutical composition comprising one of the compounds described in paragraphs 1 to 3 and a pharmaceutically acceptable carrier.

[0081] 12. The pharmaceutical composition of paragraph 11, further comprising a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (Sequence ID 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (Sequence ID 3).

[0082] 13. A kit comprising (i) one compound from any of paragraphs 1 to 3, or the composition from paragraph 11, and (ii) a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

[0083] Other embodiments are described in the following claims.

Claims

1. The following lipids 【Chemistry 1】 Alternatively, a compound consisting of a salt thereof and a CpG oligodeoxynucleotide represented by the sequence 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3' (SEQ ID NO: 1) linked at the 5' end (where X is O or S).

2. The compound according to claim 1, wherein all internucleoside groups linking the nucleoside of 5'-TCGTCGTTTTGTCGTTTTGTCGTT-3' (SEQ ID NO: 1) are phosphorothioates.

3. A composition for the treatment of cancer in human patients, comprising the compound of claim 1, wherein the treatment comprises the step of administering to a patient a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2) and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

4. The composition of claim 3, wherein the cancer is positive for human papillomavirus (HPV).

5. The composition according to claim 4, wherein the cancer is HPV type 16 positive.

6. The composition according to claim 3, wherein the cancer is head and neck squamous cell carcinoma.

7. The composition of claim 3, wherein the patient is currently receiving or has previously received platinum-containing chemotherapy.

8. The composition of claim 3, wherein the treatment involves simultaneously administering the compound and a protein having the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO:

3.

9. The composition of claim 3, wherein the treatment involves sequentially administering the compound and a protein having the amino acid sequences of SEQ ID NO: 2 and SEQ ID NO:

3.

10. A pharmaceutical composition comprising the compound of claim 1 and a pharmaceutically acceptable carrier.

11. The pharmaceutical composition of claim 10, further comprising a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

12. A kit comprising (i) the compound of claim 1, and (ii) a protein having the amino acid sequence MHQKRTAMFQ DPQERPRKLP QLCTELQTTI HDIILECVYC KQQLLRREVY DFAFRDLCIV YRDGNPYAVG DKCLKFYSKI SEYRHYCYSL YGTTLEQQYN KPLCDLLIRC INGQKPLCPE EKQRHLDKKQ RFHNGRGRWT GRCMSCCRSS RTRRETQL (SEQ ID NO: 2), and a protein having the amino acid sequence MHGDTPTLHE YMLDLQPETT DLYGYGQLND SSEEEDEIDG PAGQAEPDRA HYNIVTFCCK CDSTLRLCVQ STHVDIRTLE DLLMGTLGIV CPICSQKP (SEQ ID NO: 3).

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