Immunostimulatory micellar compositions

Abstract

The present invention relates to immunostimulatory micellar compositions, and their use in treating diseases and disorders such as cancer. In particular, the present invention relates to micellar compositions comprising a TLR7 agonist such as 1V270.

Description

Technical Field

[0001] The present invention relates to immunostimulatory micellar compositions, and their use in treating diseases and disorders such as cancer. In particular, the present invention relates to micellar compositions comprising a TLR7 agonist such as 1V270. Background Art

[0002] Toll-like receptors (TLRs) are a class of receptors expressed on various cell types and play a key role in the innate immune system. After activation, TLR activation involves the signal transduction pathway of immune activation. Several mammalian TLRs and many agonists thereof have been identified. For example, single-stranded RNA rich in guanine and uridine has been identified as the natural ligand of TLR7. In addition, several low molecular weight activators of TLR7 have been identified, including imidazoquinoline and purine-like molecules. Although TLR stimulation initiates a common signal transduction cascade (involving adapter protein MyD88, transcription factor NFκB and proinflammatory and effector cytokines), different TLRs are expressed by different cell types, however, TLR7 is mainly expressed in monocytes, plasmacytoid dendritic cells, bone marrow dendritic cells and B cells and is located in the endosomal membrane.

[0003] TLR7 has been shown to play an important role in the pathogenesis of autoimmune disorders such as systemic lupus erythematosus (SLE) and in the regulation of antiviral immunity. The TLR7 agonist Aldara (imiquimod) is an imidazoquinoline that has been approved for topical use in the treatment of warts, basal cell carcinomas, and actinic keratosis caused by papillomaviruses. Due to their ability to induce robust production of anticancer cytokines such as interleukin 12, TLR7 agonists have also been studied for cancer immunotherapy. Recent examples include delivery of TMX-202 via liposome formulations, and delivery of resiquimod via nanoparticles formed by beta-cyclodextrin.

[0004] However, repeated injections of therapeutic nanoparticle compositions have been shown to induce the antibody accelerated blood clearance (ABC) phenomenon, particularly for PEG-based liposomes, which can undermine the utility of therapeutic nanoparticle delivery systems.Therefore, there is a motivation in the art to develop more effective therapeutic nanoparticle compositions. Summary of the invention

[0005] The present inventors have surprisingly found that micellar compositions comprising a TLR7 agonist exhibit very potent anti-cancer activity without the previously recognized undesirable pharmacokinetic behavior.

[0006] In a first aspect, the present disclosure provides a micellar composition comprising: a toll-like receptor 7 (TLR7) agonist of formula (I), formula (II), formula (III), or formula (IV);

[0007]

[0008]

[0009] Where X 1 Is -O-, -S-, or –NR C ;

[0010] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0011] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or R C and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0012] R 2 each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl), halogen, nitro or cyano, or R 2 does not exist;

[0013] R a and R beach is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0014] Wherein the substituent on any alkyl, aryl or heterocyclic group is hydroxyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkylene, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 1-6 Alkoxy C 1-6 Alkylene, amino, cyano, halogen or aryl;

[0015] n is 0, 1, 2, 3, or 4;

[0016] X 2 is a bond or a linking group; and

[0017] R 3 It is a lipid;

[0018] X 3 is -N- or -CH-;

[0019] R 4 is -CH2- or -CH(R 2 )-;and

[0020] k is 0 or 1;

[0021] X 4 It is -O-, -S-, -NH-, -N(R d )-, -CH2- or -CH(R 2 )-;

[0022] R d each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -C(O)NR a R b (carbamoyl);

[0023] or a tautomer thereof;

[0024] or a pharmaceutically acceptable salt or solvate thereof, and

[0025] The ring system of formula (II) is a piperidine ring, one of the heteroatoms is a N atom and the N atom of the piperidine ring is 2 adjacent, and

[0026] wherein the purine group in any of Formulas (I), (II), (III) or (IV) undergoes tautomeric rearrangement;

[0027] and amphiphilic micelle formers.

[0028] In a second aspect, there is provided a micellar composition or a pharmaceutical composition as defined herein for use in preventing, treating or ameliorating a disease or disorder.

[0029] In a third aspect, there is provided a method for activating immune cells in a subject in vivo, comprising administering to the subject a micellar composition or a pharmaceutical composition as defined herein in an amount sufficient to activate the immune cells.

[0030] In a fourth aspect, a method for enhancing or intensifying a treatment comprising radiotherapy and / or administration of a chemotherapeutic agent or an immune checkpoint inhibitor is provided, the method comprising administering to the subject a micellar composition or a pharmaceutical composition as defined herein in combination with radiotherapy, administration of a chemotherapeutic agent and / or administration of an immune checkpoint inhibitor. BRIEF DESCRIPTION OF THE DRAWINGS

[0031] Figure 1: Figure 1A : Overview of the micelles used, micelle names, composition, molar ratio, surface charge (ζ), size, and polydispersity index (PDI). The micelles are composed of 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DOPE-PEG2000 or PEG) and the TLR7 agonist 1v270 (2,3-bis(oleoyloxy)propyl phosphate 2-(4-((6-amino-2-(2-methoxyethoxy)-8-oxo-7H-purine-9(8H)-yl)methyl)benzamido)ethyl ester) (1v270). The composition and molar ratio of each component in the micelle are shown. The micelle surface charge is expressed in mV. The micelle size is measured in nanometers (nm). The error is expressed as SEM. The micelle name is used in the figure to clarify the exact composition. Figure 1B : Micelle size distribution by number for the four micelles tested. When 1v207 was added to the formulation, the micelle size increased. Figure 1C: Micelle size distribution by number and zeta potential of the four micelles tested. When 1v207 was added to the formulation, the micelle size increased from 11.5 nm to 13-14 nm. When 1v207 was added to the formulation, the zeta potential of the micelles decreased from -4 mV to -8 mV.

[0032] Figure 2 : Structure of the TLR7 agonist 1v270 (also known as TMX-201) formulated for micellar formulation (Mw=1085.4, name: 2,3-bis(oleoyloxy)propyl 2-(4-((6-amino-2-(2-methoxyethoxy)-8-oxo-7H-purin-9(8H)-yl)methyl)benzamido)ethyl phosphate).

[0033] Figure 3: Figure 3A -B compares the antitumor studies between micelles containing 1V270 and liposomes containing 1V270. Figure 3C The antitumor activity of micellar formulation MBS8 at three doses of 50, 200 and 400 nmol per mouse was shown, with the first injection on day 9 and then 4 more injections at 4-day intervals. Figure 3D The benefit of a low dose of 50 nmol / mouse MBS8 micelles in combination with optimal PD-1 treatment is shown. Figure 3E Shown are the median survival times of the groups of mice treated with radiation therapy in combination with liposomes and micelles containing 1V270. Figure 3F The ratio of tumor to control is shown (the average tumor size of the group of mice treated with micelles or liposomes containing 1V270 in combination with radiotherapy (tumor = T) compared to the average tumor size of the control radiotherapy treated group (control = C). Calculation was performed by T / C x 100. Micelles showed significantly better antitumor activity than liposomes (p < 0.01). Figure 3G Tumor growth inhibition calculated by subtracting 100-T / C ​​ratio is shown. Micelles showed significantly better tumor growth inhibition than liposomes (p<0.01).

[0034] Figure 4: Figure 4A -G shows cytokine secretion in the plasma of mice treated with equal doses of micelles or liposomes. In general, liposomes MBS1 showed higher cytokine secretion than micelles for all measured cytokines, and MBS2 liposomes showed higher cytokine secretion than micelles for most of the tested cytokines. This suggests that micelles have a reduced risk of cytokine-related syndrome (CRS) compared to liposomes containing 1V270.

[0035] Figure 5: Figure 5A-B shows the induction of anti-PEG IgM and IgG molecules in the plasma of mice treated once with an intravenous injection of micelles or liposomes containing 1V270. Anti-PEG IgM and IgG molecules were measured by ELISA designed for this purpose. Figure 5C The specificity of the anti-PEG IgM determined using a competition assay is shown. Only PEGylated liposomes were recognized by the produced IgM, while free PEG chains, DSPE-PEG micelles and MBS8 micelles were not recognized by the anti-PEG IgM molecules.

[0036] Figure 6: Tumor growth and survival of mice bearing subcutaneous CT26 tumors and treated with 50, 100 or 200 nmol micelles in combination with radiotherapy (RT). Mice were treated on day 12 after inoculation and treated with 2Gy RT for 5 consecutive days (days 12, 13, 14, 15, 16) and micelles every 4 days for a total of 5 times (days 12, 16, 20, 24, 28). Groups contained 8-10 mice. Tumor growth curves are shown as mean ± SEM. The statistical significance of survival was determined using the Mantel-Cox test. Fig. 6A : Tumor growth in mice treated with MBS6 in combination with radiation therapy. Figure 6B : Survival rate of mice treated with MBS6 in combination with radiotherapy. Figure 6C : Tumor growth in mice treated with MBS7 in combination with radiation therapy. Fig.6D : Survival rate of mice treated with MBS7 in combination with radiation therapy. Fig. 6E : Tumor growth in mice treated with MBS8 in combination with radiation therapy. Fig. 6F : Survival rate of mice treated with MBS8 in combination with radiation therapy.

[0037] Figure 7: Comparison of the antitumor activity of MBS8 micelles containing DOPE-PEG2000 and TLR7 agonist 1v270 at a 90:10 molar ratio and administered at three levels in the CT26 model. MBS8 was administered at three doses of 50 (small black circles), 200 (grey circles) and 400 (large black circles) nmol per mouse intravenously. The two high doses of 200 and 400 nmol were significantly different from the control treatment group (p<0.001). The growth of each tumor individually was as shown in Figure 7. Figure 7B As shown, 1) control treatment (PBS), 2) low dose MBS8, 50 nmol / injection / mouse, 3) medium dose MBS8, 200 nmol / injection / mouse, and 4) high dose MBS8, 400 nmol / injection / mouse.

[0038] Figure 8: In the CT26 colon cancer syngeneic mouse tumor model, three different dose levels of MBS8 micelles were combined with αPD-1 treatment (clone RMP1-14 administered at 10 mg / kg IP). MBS8 was administered intravenously at three doses of 50, 200, and 400 nmol per mouse, with the first injection on day 9 and then 4 more injections at 4-day intervals on days 13, 17, 21, and 25. αPD-1 mAb was injected IP starting on day 11, twice a week for three weeks. The PD-1-MBS8 combination treatment was significantly stronger than PD-1 monotherapy (p<0.05, p<0.005, Wilcoxon rank sum test) ( Fig. 8A ). Figure 8B Individual tumor growth of all 10 mice in each group are shown, including 1) control (PBS treatment), 2) 50nmol of MBS8, 3) 200nmol of MBS8, 4) 400nmol of MBS8, 5) PD1 treatment alone injected at 10mg / kg IP, 6) combination therapy of PD-1 and 50nmol MBS8, 7) combination therapy of PD-1 and 200nmol MBS8, 8) combination therapy of PD-1 and 400nmol MBS8.

[0039] FIG. 9 : MBS8 enhances the anti-tumor effects of doxorubicin (AB) and doxil (CD).

[0040] Mean tumor volume ± SEM is shown, n = 10 in (A) and (C). Wilcoxon rank sum test was used for statistical data processing. Tumor volumes of individual animals of the indicated groups are shown in (B) and (D).

[0041] Figure 10: Tumor growth of mice bearing subcutaneous CT26 tumors treated with radiation therapy (RT) in combination with 100 nmol MBS8 intratumorally or 200 nmol MBS8 intravenously. When indicated, mice were treated with lipid-matched vehicle instead of MBS8. Mice were treated on day 12 after inoculation and treated with 2 Gy RT for 5 consecutive days and micelles every 4 days for a total of 5 times. Groups contained 8 mice. Tumor growth curves are shown as mean ± SEM. Fig. 10A : Tumor growth in mice treated with MBS8 intratumorally or intravenously in combination with radiation therapy. Fig. 10B : Tumor growth in mice treated with vehicle intratumorally or intravenously in combination with radiation therapy. Fig. 10C : Shown is the survival rate of mice treated with MBS8 by intratumoral route, either micelles without TLR7 agonist (vehicle) or MBS8 with TLR7 agonist 1V270. Shown are groups combined with or without radiation therapy. Fig. 10D: Shows the survival rate of mice treated with MBS8 by intravenous route, whether micelles without TLR7 agonist (vehicle) or MBS8 with TLR7 agonist 1V270. Shows groups combined with or without radiation therapy. The group treated with RT+intravenous MBS8 showed significantly better antitumor activity than RT alone (p=0.015), while the group treated with RT+intratumoral MBS8 did not show significantly better antitumor activity than RT alone, n=8 / group.

[0042] Figure 11: Tumor growth and survival of mice bearing subcutaneous EL4 or MC38 tumors treated with radiation therapy (RT) in combination with 200 nmol MBS8 or vehicle. Tumor growth curves are shown as mean ± SEM.

[0043] EL4-bearing mice were treated starting at day 7 post-inoculation and were treated with 2 Gy RT for 3 consecutive days and with MBS8 or vehicle every 4 days for a total of 5 treatments. Groups contained 9-10 mice. Fig.11A : Tumor growth in treated EL4-bearing mice. Fig. 11B : Survival rate of treated EL4-bearing mice.

[0044] MC38-bearing mice were treated starting at day 10 post-inoculation and were treated with 2 Gy RT for 5 consecutive days and MBS8 or vehicle every 2 days (q2d), 4 days (q4d), 7 days (q7d) for a total of 5 treatments. Groups contained 9 mice. Fig. 11C : Tumor growth in treated MC38-bearing mice. Fig.11D : Survival rate of treated MC38-bearing mice.

[0045] Figure 12: MBS8 administration in cynomolgus monkeys. (A) C-reactive protein (CRP) was measured immediately before drug administration and 8, 24 and 72 hours after infusion. Data represent mean values ​​(n=3). Arrows indicate the peak CRP level 24 hours after each injection. The corresponding doses are shown in Table 1. Fig. 12B Indicated. (B) Body temperature was monitored daily. Mean ± SD values ​​are shown (n = 3). The drug administration days and corresponding doses are indicated. The dotted line indicates the normal range based on baseline measurements during the adaptation period (day -22 to day 0).

[0046] Figure 13 is an in vivo efficacy study using 12 syngeneic subcutaneous tumor models. (A) Table summarizes the results of all 12 models. When the average tumor size reached about 100 mm 3At the time of treatment, mice were randomized and treated with the indicated treatment measures. PD-1 (clone RMP1-14) was administered intraperitoneally at 10 mg / kg, 6 times q4d, and MBS8 was administered intravenously at 300 μg / mouse, 5 times q4d. The same dosing regimen was used for the combination study. All groups included 10 mice / group. TGI, tumor growth inhibition; CR, complete responder; NA, not applicable. (BD) Effect of MBS8 on subcutaneous tumor growth. Upper figure: Tumor growth curves in the indicated animal groups. Mean tumor volume + SEM is shown. Significant two-sided p-values ​​(Wilcoxon rank sum test) are shown. Middle and lower figures: Tumor growth curves of individual animals in the indicated treatment groups are shown. (B) EMT-6, (C) Hepa 1-6 and (D) Pan02 represent three different response patterns found in 12 tumor groups. CR, complete responder; RR, re-challenge resistance; * Two animals died at the start of treatment and were excluded from statistical analysis. (E) Rejection of re-challenge EMT-6 tumors. Mice treated with MBS8 (n = 8) or MBS8 + anti-PD-1 (n = 10) in the EMT-6 model showed CR and tumor-free mice for at least three weeks were re-challenge with EMT-6 cells in the contralateral flank and tumor growth was monitored for 29 days. Naive mice (n = 5) inoculated with EMT-6 cells were used as untreated controls. (F) Immune memory response in mice treated with MBS8 alone or in combination with α-PD-1. CR from the CT26 model was re-challenge, and all showed rejection of re-challenge tumors. In these animals, ELISPOT was used to assess the presence of tumor-specific T cells. Splenocytes from untreated CT26 tumor-bearing mice (control), mice previously treated with MBS8 and resistant to rechallenge (single), mice treated with the MBS8+α-PD-1 combination (combination), and naive tumor-free mice stimulated in vitro with (+AH-1) or without the CT26 tumor-specific antigen AH-1 (naive) were analyzed. The number of cells expressing IFN-γ / well was quantified. Mean + SEM is shown; n = 5 mice / group. Statistical analysis was performed using the Wilcoxon rank sum test. Significant two-sided p-values ​​are shown.

[0047] Figure 14 is a comparative efficacy of MBS8 administered at different dose regimens. The CT26 subcutaneous model was used. When the average tumor size reached approximately 100 mm 3 At 4 hr, mice were randomized and treated intravenously with 200 nmol / mouse at the indicated dose regimens. (A) Mean tumor volume + SEM is shown. Significant two-sided p-values ​​(two-tailed Wicoxon rank sum test) are shown. (B) Tumor growth curves for individual animals. Animals with tumors <40 mm3 at the last measurement and showing continuous volume reduction were considered complete responders (CR).

[0048] Figure 15: MBS8 and R848 treatment of mice bearing subcutaneous CT26 tumors. Mice bearing CT26 tumors were treated starting on day 11 after inoculation and were treated intravenously with MBS8 (100, 200 or 300 nmol) or R848 (200 nmol) on a q4d schedule for a total of 5 treatments. n = 9 mice / group. (A) Mean tumor growth curves ± SEM are shown. (B) Survival rate of treated CT26-bearing mice. The statistical significance of survival was determined using the Mantel-Cox test, ****p<0.0001, *p<0.05, (C) Weight change relative to baseline, mean weight ± SEM is shown.

[0049] Fig.16 : Cell types in tumors of mice treated with MBS8 monotherapy based on gene expression analysis. Mice were treated with 200 nmol MBS8 on a q4d schedule, with 1 or 3 injections. Tumors were collected on day 0 (untreated), 1 day after the first injection, 2 days after the first injection, 4 days after the first injection, 14 days after the first injection, 2 days after the third injection, and 4 days after the third injection, for all groups, n=3-6. Gene expression analysis was performed on the extracted RNA, and the data were analyzed for cell types using the Nanostring Advanced Software Module. A two-way ANOVA with multiple comparison correction was performed to compare different time points with untreated. * indicates p<0.05; ** indicates p<0.01; *** indicates p<0.001 and **** indicates p<0.0001

[0050] Figure 17: Effect on the tumor microenvironment shortly after injection of MBS8 based on flow cytometry analysis. CT26-bearing mice were injected with 200 nmol MBS8 and tumors were evaluated by flow cytometry. Tumors were collected 1, 3, and 6 hours after injection. All figures are shown as mean ± SEM. Statistical significance compared to untreated tumors was determined using the Kruskal-Wallis test and Dunn's multiple comparison test, where * indicates p < 0.05; ** indicates p < 0.01; and *** indicates p < 0.001. n = 3-4. MFI = median fluorescence intensity.

[0051] Figure 18: Effects on the tumor microenvironment during treatment based on flow cytometry analysis. Mice bearing CT26 tumors were treated with 200 nmol MBS6 every 4 days on day 15 after cancer cell inoculation, and tumors and spleens were analyzed two days after the second injection. All figures are shown as mean ± SEM. Statistical significance compared to untreated tumors was determined using the Mann-Whitney U test, where * indicates p < 0.05; ** indicates p < 0.01; n = 5.

[0052] Fig.19 : Dosing schedule study of Doxil-treated CT26 tumor-bearing mice on days 9, 13, and 17 for all groups (black circles), MBS8 was additionally administered to the Doxil+MBS8 pre-dose+post-dose treatment group on days 9 and 19, 23, 27, and 31, and MBS8 was additionally administered to the Doxil+MBS8 post-dose treatment group on days 19, 23, 27, and 31. All figures are shown as mean ± SEM. Statistical significance of comparisons between the indicated groups was determined using the Mann-Whitney U test at the indicated significance level, n = 10.

[0053] Figure 20: Mean (+ / -SD) plasma concentration-time profiles of MBS8 in female and male rats on day 1 (first dose) and day 15 (one week after the last of two doses separated by 1 week) after administration of 3 mg / kg / day MBS8 (1V270) by intravenous infusion over 1 hour on a linear scale (A) and a semi-logarithmic scale (B). The figure demonstrates the indifferent kinetics of MBS8 administered after 1 day and 15 days (third dose), respectively. This observation supports that the micelles of the present invention do not induce accelerated blood clearance (ABC).

[0054] Figure 21: Overlay of individual plasma concentration-time curves of 1V270 in 1 female and 1 male cynomolgus monkeys on day 1 (A) (first dose) and day 15 (B) (one week after the last dose of two doses separated by 1 week) after administration of 0.3, 1 and 3 mg / kg / day MBS8 (1V270) by intravenous infusion over 1 hour on a linear scale. The figure shows no difference in kinetics of MBS8 administered after 1 day (first dose) and 15 days (third dose after weekly dosing), respectively. This observation supports that the micelles of the present invention do not induce accelerated blood clearance (ABC). DETAILED DESCRIPTION

[0055] definition

[0056] As disclosed herein, "lipid" refers to a group of substances that contain at least one hydrophobic portion and are insoluble in water. An exemplary group of lipids can be, but is not limited to, fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids, prenol lipids, glycolipids, and polyketides.

[0057] As used herein, the term "prevention" refers to preventing a disease or preventing the spread of a disease.

[0058] The term "treatment" as used herein refers to combating a disease or disorder. "Treatment" as used herein includes any desired effect on the symptoms or pathology of a disease or condition as described herein, and may even include minimal changes or improvements in one or more measurable markers of the disease or condition being treated. "Treatment" or "treating" does not necessarily indicate complete eradication or cure of a disease or condition or its associated symptoms.

[0059] The term "PEG" as used herein refers to polyethylene glycol.

[0060] As used herein, the term "amphiphilic micelle former" refers to an agent that is "amphiphilic," i.e., has both hydrophilic (water-loving, polar) and lipophilic (fat-loving) properties while also being able to form micelles. The amphiphilic micelle formers disclosed herein are capable of forming micelles in a mixture with other compounds in solution.

[0061] As used herein, the term "linking group" refers to a group of bonding atoms, such as a functional group, that covalently connects at least two parts of a molecule. In one example, the linking group is a carbonyl group, i.e., "-C(O)-", which can bind, for example, an amine "H2N-alkyl(1)" at one end and an alkyl(2) at the other end, thereby forming an amide "alkyl(2)-C(O)-NH-alkyl(1)".

[0062] The terms “anti-PD-1,” “α-PD-1,” and “a-PD-1” are used interchangeably herein to refer to the group of immune checkpoint inhibitors, anti-PD-1.

[0063] The terms “anti-PD-L1,” “α-PD-L1,” and “a-PD-L1” are used interchangeably herein to refer to the group of immune checkpoint inhibitors, anti-PD-L1.

[0064] As used herein, the term "MBS6" refers to a micellar composition of DSPE-PEG2000:1V270 at a molar ratio of 80:20.

[0065] The term "MBS7" as used herein refers to a micellar composition of DSPE-PEG2000:1V270 at a molar ratio of 95:5.

[0066] The term "MBS8" as used herein refers to a micellar composition of DSPE-PEG2000:1V270 at a molar ratio of 90:10.

[0067] Micelles

[0068] Micelles are aggregated particles of amphiphilic molecules dispersed in a liquid colloid. Most micelles in aqueous solution form aggregated particles with their hydrophilic head groups in contact with the surrounding hydrophilic solvent, thereby isolating the hydrophobic tail region in the center of the micelle.

[0069] In the compound forming micelles, micelles made of polyethylene glycol-phosphatidylethanolamine (PEG-PE) have particularly attractive characteristics, such as good stability, long life and can be accumulated in areas with abnormal vasculature (entering areas with leaky vasculature, such as tumors) via enhanced permeability and retention. In addition, these micelles can be "targeted" by attaching specific targeting ligand molecules to the micelle surface, or can be composed of stimulus-responsive amphiphilic block copolymers. Adding a second component such as a surfactant or another hydrophobic material to the main micelle forming material further improves the solubilization ability of micelles without compromising their stability. Compared with other nanoparticles such as liposomes, micelles are easier to prepare and can be prepared by lipid mixtures and ultrasonic treatment. The size of micelles can be determined by various techniques known to those skilled in the art. Dynamic light scattering (DLS) experiments can be performed using, for example, a Malvern Zetasizer Nano ZS instrument suitable for measuring the size and size distribution of micelles formed in aqueous solution. The diameter measured in DLS refers to the value of how particles diffuse in the fluid, and is referred to as the hydrodynamic diameter.

[0070] The diameters of the micelles disclosed herein are expressed as numerical averages.

[0071] In one embodiment, the diameter of the micelle disclosed herein is between 5 nm and 50 nm, such as between 6 nm and 46 nm, such as between 7 nm and 42 nm, such as between 8 nm and 38 nm, such as between 9 nm and 34 nm, such as between 10 nm and 34 nm, such as between 11 nm and 30 nm, such as between 12 nm and 26 nm.

[0072] In one embodiment, the diameter of the micelle is from 5 nm to 39 nm, such as from 5 nm to 20 nm, such as from 20 nm to 30 nm, such as from 30 nm to 35 nm, such as from 35 nm to 39 nm.

[0073] In one embodiment, the diameter of the micelle is 5nm to 39nm, such as 5nm to 38nm, such as 5nm to 37nm, such as 5nm to 36nm, such as 5nm to 35nm, such as 5nm to 34nm, such as 5nm to 33nm, such as 5nm to 32nm, such as 5nm to 31nm.

[0074] In one embodiment, the diameter of the micelle is between 5nm and 25nm, such as between 6nm and 24nm, such as between 7nm and 23nm, such as between 8nm and 22nm, such as between 9nm and 21nm, such as between 10nm and 20nm, such as between 11nm and 19nm, such as between 12nm and 18nm, such as between 13nm and 17nm, such as between 14nm and 16nm, such as 15nm.

[0075] The "amphiphilic micelle forming agent" of the present disclosure may comprise a phospholipid in some embodiments. The structure of a phospholipid generally comprises two hydrophobic fatty acid "tails" and a hydrophilic "head" group comprising a phosphate group. These two components are linked together by a glycerol molecule. The phosphate group may be modified with simple organic molecules such as choline, ethanolamine or serine. In some embodiments, simple organic molecules serve as linking groups for polymers such as PEG.

[0076] In one embodiment, the amphiphilic micelle former is selected from the group consisting of poloxamers, poloxamines, PEG-polyesters, PEG-polyanhydrides, PEG-polyamino acids, phospholipids, polysorbates, and polyoxyethylene alkyl ethers.

[0077] In one embodiment, the PEG-polyester is selected from the group consisting of: PEG-poly(lactic acid) (PEG-PLA), PEG-poly(lactic-co-glycolic acid) (PLGA), and PEG-poly(ε-caprolactone) (PCL).

[0078] In one embodiment, the PEG-polyanhydride is PEG-polysebacic anhydride (PSA).

[0079] In one embodiment, a micellar composition comprising PEG-polyamino acids is provided, wherein the PEG-polyamino acids are selected from: PEG-poly (L-histidine), PEG-poly (L-aspartic acid), PEG-poly (L-asparagine), PEG-poly (L-glutamic acid), PEG-poly (L-glutamine) and PEG-poly (L-lysine).

[0080] In one embodiment, the amphiphilic micelle former is a phospholipid conjugated to polyethylene glycol (PEG). In one embodiment, the phospholipid conjugated to PEG is conjugated via a carbonyl group.

[0081] An example of a preferred amphiphilic micelle former is DSPE-PEG2000, exemplified below as an ammonium salt:

[0082]

[0083] (DSPE-PEG2000)

[0084] DSPE-PEG2000 comprises phosphatidylethanolamine bound to PEG via a carbonyl group. Phosphatidylethanolamine comprises glycerol esterified with two fatty acids and phosphate. While the phosphate group is bound to the choline in phosphatidylcholine, it is bound to the ethanolamine in phosphatidylethanolamine. The two fatty acids can be the same or different and are usually in the 1,2 position, but can also be in the 1,3 position. In some embodiments, the termini of the PEG are aminated, i.e., bound to NH2.

[0085] Polyethylene glycol (PEG)

[0086] In one embodiment, the micelles of the present disclosure comprise PEG. In one embodiment, the PEG is in the form of PEG conjugated to a phospholipid. The size of the PEG is between PEG350 and PEG30.000.

[0087] In one embodiment, the size of the PEG is between PEG350 and PEG5000, such as between PEG550 and PEG4000, such as between PEG750 and PEG3000, such as between PEG1000 and PEG3000, preferably the size of the PEG is PEG2000.

[0088] Phospholipids of the present disclosure

[0089] The phospholipids disclosed herein can be part of a TLR7 agonist or an amphiphilic micelle former. Examples of phospholipids disclosed in the context of an amphiphilic micelle former can also be used as part of a TLR7 agonist, and vice versa.

[0090] In one embodiment, a micellar composition according to the present disclosure is provided, which comprises a phospholipid, wherein the phospholipid comprises one or more alkyl chains, and the one or more alkyl chains are one or more C8-C24 alkyl chains, such as C10-C22, such as C12-C20, preferably C14-C18, most preferably C16-C18 saturated alkyl chains or unsaturated alkyl chains.

[0091] In one embodiment, a micellar composition according to the present disclosure is provided, which comprises a phospholipid, wherein the phospholipid comprises phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidic acid (PA), diphosphatidylglycerol (DPG) or a phosphatidyl alcohol.

[0092] In one embodiment, the phosphatidylethanolamine is selected from 1,2-dioleoyl-phosphatidylethanolamine, 1,2-dipalmitoyl-phosphatidylethanolamine, 1,2-dimyristoyl-phosphatidylethanolamine, 1,2-distearoyl-phosphatidylethanolamine, 1-oleoyl-2-palmitoyl-phosphatidylethanolamine, 1-oleoyl-2-stearoyl-phosphatidylethanolamine, 1-palmitoyl-2-oleoyl-phosphatidylethanolamine and 1-stearoyl-2-oleoyl-phosphatidylethanolamine.

[0093] In one embodiment, the phospholipid conjugated to PEG is selected from: 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)-PEG, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE)-PEG and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE)-PEG.

[0094] In one embodiment, a micellar composition comprising a phospholipid conjugated to PEG is provided, wherein the phospholipid is: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG.

[0095] In one embodiment, the amphiphilic micelle former is DSPE-PEG2000.

[0096] Pharmaceutical composition

[0097] The micelles of the present disclosure may be used as a component of a pharmaceutical formulation. Thus, in one embodiment, the micellar composition of the present disclosure is a pharmaceutical composition. In one embodiment, a pharmaceutical composition comprising a micellar composition as defined herein is provided.

[0098] Any form of such formulations suitable for administration to a mammal is contemplated.

[0099] The pharmaceutical formulation according to the present disclosure is preferably in the form of a solution, dispersion, suspension, lyophilisate or frozen.

[0100] In one embodiment, the route of administration can be intravenous, intratumoral, oral, subcutaneous, intradermal, intramuscular, intranasal, intraperitoneal, pulmonary or renal administration.

[0101] In one embodiment, the micellar composition according to the present disclosure comprises an amphiphilic micelle former and a TLR7 agonist, wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 50:50 to 99.5:0.5, such as 60:40 to 99:1, such as 70:30 to 98:2, such as 80:20 to 95:5, for example 95:5, 90:10 or 80:20.

[0102] In one embodiment, a micellar composition as disclosed herein is provided, wherein the composition comprises a TLR7 agonist at a molar concentration between 1% and 25%, such as 1%, such as 2%, such as 3%, such as 4%, such as 5%, such as 6%, such as 7%, such as 8%, such as 9%, such as 10%, such as 11%, such as 12%, such as 13%, such as 14%, such as 15%, such as 16%, such as 17%, such as 18%, such as 19%, such as 20%, such as 21%, such as 22%, such as 23%, such as 24%, such as 25%.

[0103] There is further provided a micellar composition or a pharmaceutical composition as defined herein for use in preventing, treating or ameliorating a disease or disorder.

[0104] Therapeutic uses and methods

[0105] The micellar composition of the present disclosure can be used to prevent, treat or ameliorate cancer, infectious diseases, inflammatory conditions or diseases, autoimmune diseases or allergies. In one embodiment, the micellar composition of the present disclosure is used for cancer treatment.

[0106] Thus, the micellar composition or pharmaceutical composition can be used to treat cancer; infectious diseases; inflammatory conditions or diseases; autoimmune diseases; or allergies.

[0107] In one embodiment, the disease or disorder is cancer, such as colon cancer. In one embodiment, the disease or disorder is cancer, such as a solid tumor.

[0108] Infectious Diseases

[0109] Another aspect of the present disclosure is to provide prevention or treatment of infectious diseases by administering the micellar composition disclosed herein to a subject. In a preferred embodiment, the micellar composition for preventing or treating infectious diseases is MBS8. The micellar composition disclosed herein can promote the prevention or treatment of infectious diseases in humans and livestock. In one embodiment, the infectious disease is a viral infection or a bacterial infection. In a preferred embodiment, the treatment of infectious diseases is preventive. Therefore, in one embodiment, a method for preventing infectious diseases is provided by administering a micellar composition such as MBS8 to a subject in need. Suitable subjects for preventive treatment may be, but are not limited to, healthcare professionals and / or other people who are in close contact with infected subjects. These suitable subjects have an increased risk of acquiring infection, and therefore, preventive treatment by the micellar composition disclosed herein is advantageous.

[0110] Combination therapy

[0111] In one embodiment, the treatment of cancer is enhanced by a combination of existing treatments such as monoclonal antibodies (trastuzumab, rituximab, cetuximab), radiotherapy, chemotherapy, or immune checkpoint inhibitors such as pembrolizumab, ipilimumab. Therefore, in one embodiment, the treatment of cancer is a combination therapy, which further includes administering a monoclonal antibody to a subject with cancer.

[0112] As demonstrated in Examples 11, 13, 14, 15, 16, 18 and 24, when the micellar composition or pharmaceutical composition as disclosed herein is administered in combination with radiation therapy, chemotherapeutic agents or immune checkpoint inhibitors, an enhanced or synergistic effect can be obtained. Thus, in one embodiment, the treatment of cancer is a combination therapy further comprising radiation therapy.

[0113] Certain types of chemotherapy are particularly relevant to the combination of TLR7 valences; These are chemotherapeutic compounds that induce so-called "immunogenic cell death" (ICD). As shown in Examples 14 and 24, micellar compositions comprising 1V270 significantly enhance the efficacy of doxorubicin and doxil and result in effective treatment. In one embodiment, the treatment of cancer is a combination therapy, which further includes the administration of chemotherapeutic agents such as doxorubicin or doxil. In one embodiment, the chemotherapeutic agent is selected from doxorubicin, Doxil, epirubicin, cyclophosphamide, bortezomib and oxaliplatin. In one embodiment, the treatment of cancer is a combination therapy, which further includes the administration of immune checkpoint inhibitors, such as monoclonal antibodies targeting PD-1, PD-L1 or CTLA-4, such as α-PD-1, α-PD-L1 or α-CTLA-4, for example, atezolizumab, avelumab, durvalumab, nivolumab, tisellizumab (Tisellizumab), pembrolizumab or ipilimumab. Preferably, the immune checkpoint inhibitor is α-PD-1, such as nivolumab or pembrolizumab. In a preferred embodiment, the micellar composition is MBS8 and the immune checkpoint inhibitor is nivolumab or pembrolizumab.

[0114] As demonstrated in Example 18, the micellar composition is extremely effective in treating various cancers. This effect is demonstrated as a monotherapy and in combination with α-PD-1, even in the treatment of cancers that do not respond to α-PD-1 monotherapy. In one embodiment, the combination therapy comprises administering a micellar composition and α-PD-1 to treat cancer, in particular a cancer selected from the group consisting of liver cancer, pancreatic cancer, lymphoma, breast cancer, and colon cancer. In one embodiment, the combination therapy comprises administering a micellar composition and α-PD-1 for the treatment of a cancer selected from the group consisting of prostate cancer and kidney cancer. In particular, the combination of micellar compositions and α-PD-1 is effective in treating cancers that do not respond to α-PD-1 monotherapy.

[0115] In one embodiment, the micellar composition used in the combination therapy is MBS8. The preferred combination therapy is MBS8 and α-PD-1. In particular, the preferred combination therapy is effective in treating cancers that do not respond to anti-PD-1 monotherapy.

[0116] As shown in Example 13, the combination of micellar compositions comprising 1V270 and α-PD-1 was highly potent, resulting in complete remission in at least 90% of treated mice bearing the CT26 model. In one embodiment, MBS8 is administered in combination with a therapeutic antibody targeting the PD-1 / PD-L1 pathway to a cancer patient in need of treatment. In one embodiment, the therapeutic antibody targeting the PD-1 / PD-L1 pathway is selected from the group consisting of: atezolizumab, avelumab, durvalumab, nivolumab, pembrolizumab, spartalizumab / PDR001, tislelizumab, BCD-100, TSR-042, carrelizumab, IBI308, KNO35, and CS1001.

[0117] Combining monoclonal antibodies with TLR7 agonists is beneficial by activating complement dependent cytotoxicity (CDC), antibody dependent cellular cytotoxicity (ADCC), and antibody dependent cellular phagocytosis (ADCP). This is relevant for antibodies against, for example, CD20, EGFR, CD38, and HER2. ADCC, ADCP, and CDC-mediated tumor cell killing depends on the ability of the micelle composition ( Fig.16 ), especially MBS8-activated NK cells, macrophages and neutrophils.

[0118] In one embodiment, MBS8 is administered to a cancer patient in combination with a monoclonal antibody targeting CD20.

[0119] In one embodiment, MBS8 is administered to a cancer patient in combination with a monoclonal antibody targeting epidermal growth factor receptor (EGFR).

[0120] In one embodiment, MBS8 is administered to a cancer patient in combination with a monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2).

[0121] In one embodiment, MBS8 is administered to a cancer patient in combination with a monoclonal antibody targeting CD38. In a specific embodiment, the monoclonal antibody targeting CD38 is selected from daratumumab and isatuximab.

[0122] In one embodiment, MBS8 is administered to a cancer patient in combination with a monoclonal antibody selected from the group consisting of: utuximab, obinutuzumab, ofatumumab, ibritumomab tiuxetan, rituximab, tositumomab, martin-dietuximab, necituzumab, panitumumab, cetuximab, trastuzumab, trastuzumab-dkst, trastuzumab emtansine, BAT8001, pertuzumab, magetuximab, trastuzumab deruxtecan, trastuzumab duocarmazine, daratumumab, and isatuximab.

[0123] In one embodiment, MBS8 is administered in combination with an antibody targeting CD47, such as mololimab.

[0124] In one embodiment, the treatment of cancer is performed as a monotherapy comprising the administration of a micellar composition or a pharmaceutical composition as disclosed herein.

[0125] As used herein, the term "prevention" refers to preventing a disease or preventing the spread of a disease.

[0126] The term "treatment" as used herein refers to combating a disease or disorder. "Treatment" as used herein includes any desired effect on the symptoms or pathology of a disease or condition as described herein, and may even include minimal changes or improvements in one or more measurable markers of the disease or condition being treated. "Treatment" or "treating" does not necessarily indicate complete eradication or cure of a disease or condition or its associated symptoms.

[0127] As used herein, the term "amelioration" refers to a reduction in the severity of the symptoms of a disease or condition. Improvement of a patient's condition, or activities that attempt to correct or at least make intolerable symptoms associated with a patient's condition more tolerable, is considered "ameliorative" treatment.

[0128] In one embodiment, the micellar composition is used to prevent, treat or ameliorate cancer.

[0129] In one embodiment, the micellar composition is used to prevent, treat or ameliorate infectious diseases.

[0130] In one embodiment, a method for activating immune cells in a subject in vivo is provided, comprising administering to the subject a micellar composition or a pharmaceutical composition as defined herein in an amount sufficient to activate the immune cells. A preferred subject is a human, such as a human suffering from cancer.

[0131] TLR7 agonists

[0132] Toll-like receptor 7 (also known as TLR7) is a protein encoded by the TLR7 gene in humans. It is a member of the toll-like receptor (TLR) family and plays an important role in pathogen recognition and innate immune activation. TLR7 agonists have been studied for cancer immunotherapy due to their ability to induce robust production of anticancer cytokines such as interleukin-12.

[0133] The micellar composition of the present disclosure comprises: a toll-like receptor 7 (TLR7) agonist of formula (I), formula (II), formula (III), or formula (IV);

[0134]

[0135] Where X 1 Is -O-, -S-, or –NR C ;

[0136] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0137] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or R C and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0138] R 2 each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl), halogen, nitro or cyano, or R 2 does not exist;

[0139] R a and R b each is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0140] Wherein the substituent on any alkyl, aryl or heterocyclic group is hydroxyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkylene, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 1-6 Alkoxy C 1-6 Alkylene, amino, cyano, halogen or aryl;

[0141] n is 0, 1, 2, 3, or 4;

[0142] X 2 is a bond or a linking group; and

[0143] R 3 It is a lipid;

[0144] X 3 is -N- or -CH-;

[0145] R 4 is -CH2- or -CH(R 2 )-;and

[0146] k is 0 or 1;

[0147] X 4 It is -O-, -S-, -NH-, -N(R d )-, -CH2- or -CH(R 2 )-;

[0148] R d each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -C(O)NRa R b (carbamoyl);

[0149] or a tautomer thereof;

[0150] or a pharmaceutically acceptable salt or solvate thereof, and

[0151] The ring system of formula (II) is a piperidine ring, one of the heteroatoms is a N atom and the N atom of the piperidine ring is 2 adjacent, and

[0152] wherein the purine group in any of Formulas (I), (II), (III) or (IV) undergoes tautomeric rearrangement;

[0153] and amphiphilic micelle formers.

[0154] It will be appreciated that the purine group in any of Formula (I), (II), (III) or (IV) undergoes tautomeric rearrangements.

[0155] In one embodiment, there is provided a micellar composition according to the present disclosure, wherein the TLR7 agonist has formula (I):

[0156]

[0157] Where X 1 Is -O-, -S-, or –NR C ;

[0158] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0159] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or R C and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0160] R 2 each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl), halogen, nitro or cyano, or R 2 does not exist;

[0161] R a and R b each is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0162] Wherein the substituent on any alkyl, aryl or heterocyclic group is hydroxyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkylene, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 1-6 Alkoxy C 1-6 Alkylene, amino, cyano, halogen or aryl;

[0163] n is 0, 1, or 2;

[0164] X 2 is a bond or a linking group; and

[0165] R 3 It is a lipid;

[0166] or a tautomer thereof;

[0167] or a pharmaceutically acceptable salt or solvate thereof, and

[0168] wherein the purine group undergoes tautomeric rearrangement.

[0169] In one embodiment, there is provided a micellar composition according to the present disclosure, wherein the TLR7 agonist has formula (I):

[0170]

[0171] Where X 1 Is -O-, -S-, or –NRC ;

[0172] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0173] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or R C and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0174] R 2 does not exist;

[0175] n is 0;

[0176] X 2 is a bond or a linking group; and

[0177] R 3 It is a lipid;

[0178] or a tautomer thereof;

[0179] or a pharmaceutically acceptable salt or solvate thereof, and

[0180] wherein the purine group undergoes tautomeric rearrangement. It will be appreciated by those skilled in the art that when R 2 When it does not exist, n must be 0, that is, it does not exist.

[0181] In one embodiment, there is provided a micellar composition according to the present disclosure, wherein the TLR7 agonist has formula (I):

[0182]

[0183] Where X 1 is-O-;

[0184] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0185] R2 does not exist;

[0186] n is 0;

[0187] X 2 is a bond or a linking group; and

[0188] R 3 It is a lipid;

[0189] or a tautomer thereof;

[0190] or a pharmaceutically acceptable salt or solvate thereof, and

[0191] wherein the purine group undergoes tautomeric rearrangement.

[0192] In one embodiment, there is provided a micellar composition according to the present disclosure, wherein the TLR7 agonist has formula (V):

[0193]

[0194] Where X 1 is-O-;

[0195] R 1 is hydrogen, (C1-C6) alkyl, substituted (C1-C6) alkyl, C6 aryl or substituted C6 aryl, C 5-6 Heterocyclic, substituted C 5-6 Heterocyclic ring;

[0196] X 2 is a bond or a linking group; and

[0197] R 3 It is a lipid;

[0198] or a tautomer thereof;

[0199] or a pharmaceutically acceptable salt or solvate thereof, and

[0200] wherein the purine group undergoes tautomeric rearrangement.

[0201] In one embodiment, there is provided a micellar composition according to the present disclosure, wherein the TLR7 agonist has formula (VI):

[0202]

[0203] Where X 1 is-O-;

[0204] R 1 is hydrogen, (C1-C6) alkyl, substituted (C1-C6) alkyl, C6 aryl or substituted C6 aryl, C 5-6Heterocyclic, substituted C 5-6 Heterocyclic ring;

[0205] X 2 is a bond or a linking group; and

[0206] R 3 It is a lipid;

[0207] or a tautomer thereof;

[0208] or a pharmaceutically acceptable salt or solvate thereof, and

[0209] wherein the purine group undergoes tautomeric rearrangement.

[0210] In one embodiment, a micellar composition according to the present disclosure is provided, wherein the TLR7 agonist has formula (VII):

[0211]

[0212] Where X 2 is a bond or a linking group; and

[0213] R 3 It is a lipid;

[0214] or a tautomer thereof;

[0215] or a pharmaceutically acceptable salt or solvate thereof, and

[0216] wherein the purine group undergoes tautomeric rearrangement.

[0217] In one embodiment, a micellar composition according to the present disclosure is provided, wherein the TLR7 agonist has formula (VIII):

[0218]

[0219] Where R 3 It is a lipid;

[0220] or a tautomer thereof;

[0221] or a pharmaceutically acceptable salt or solvate thereof, and

[0222] wherein the purine group undergoes tautomeric rearrangement.

[0223] In one embodiment, X 2Selected from: a bond, -O-, -C(O)-(carbonyl), (C1-C6)alkyl, substituted (C1-C6)alkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, -C(O)-(C1-C6)alkyl (alkanoyl), substituted -C(O)-(C1-C6)alkyl, -C(O)-(C1-C6)alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl); and

[0224] R a and R b each is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0225] And among them

[0226] X 1 Is -O-, -S- or -NR C ;

[0227] R 1 is hydrogen, C1-C6 alkyl, or substituted (C1-C6) alkyl;

[0228] R C It is hydrogen, C 1-6 Alkyl or substituted C 1-6 Alkyl; or R C and R 1 Together with the nitrogen to which they are attached they form a heterocyclic or substituted heterocyclic ring.

[0229] In a preferred embodiment, X 2 In a preferred embodiment, according to the formula disclosed herein, X 2 is -C(O)-(carbonyl) and R 3 It is 1,2-dioleoyl-phosphatidylethanolamine.

[0230] In one embodiment, R 3is a lipid selected from the group consisting of: phospholipids containing one or two carboxylic acid esters; steranes, such as cholesterol; glycolipids; and glycerides. 3 It is a phospholipid containing one or two carboxylic acid esters.

[0231] As disclosed herein, "lipid" refers to a group of substances that contain at least one hydrophobic portion and are insoluble in water. An exemplary group of lipids can be, but is not limited to, fatty acids, glycerolipids, glycerophospholipids, sphingolipids, sterol lipids, prenol lipids, glycolipids, and polyketides.

[0232] In a particularly preferred embodiment, the TLR7 agonist according to formula (I) has a structure according to formula (IA), wherein formula (I) is defined as follows:

[0233] X 1 is-O-;

[0234] R 1 2-methoxy-1-ethyl

[0235] R 2 does not exist;

[0236] X 2 is a carbonyl group; and

[0237] R 3 is 1,2-dioleoyl- sn -glycero-3-phosphoethanolamine (DOPE);

[0238] or a pharmaceutically acceptable salt or solvate thereof;

[0239]

[0240] The compound according to formula (IA) is also known in the literature as 1V270, from eg US 8,357,374.

[0241] Thus, in a preferred embodiment, a micellar composition is provided, wherein the TLR7 agonist has a structure according to formula (IA):

[0242] or a tautomer thereof;

[0243] The halogen atom mentioned herein refers to a fluorine atom, a chlorine atom, a bromine atom or an iodine atom.

[0244] Aryl refers to C6-10 monocyclic or condensed ring aromatic groups, such as phenyl, indenyl or naphthyl.

[0245] Heterocyclic or heterocycle (Het) refers to a monocyclic saturated heterocyclic group or an unsaturated monocyclic or fused heterocyclic group containing at least one heteroatom, such as 0-3 nitrogen atoms, 0-1 oxygen atoms (-O-) and 0-1 sulfur atoms (-S-). Non-limiting examples of saturated monocyclic heterocyclic groups include 5 or 6-membered saturated heterocyclic groups, such as tetrahydrofuranyl, pyrrolidinyl, morpholinyl, piperidinyl, piperazinyl or pyrazolidinyl. Non-limiting examples of unsaturated monocyclic heterocyclic groups include 5 or 6-membered unsaturated heterocyclic groups, such as furanyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, thienyl, pyridinyl or pyrimidinyl. Non-limiting examples of unsaturated fused heterocyclic groups include unsaturated bicyclic heterocyclic groups, such as indolyl, isoindolyl, quinolyl, benzothiazolyl, chromanyl, benzofuranyl, etc. The Het group may be a saturated heterocyclic group or an unsaturated heterocyclic group, such as a heteroaryl group.

[0246] Non-limiting examples of heterocycles include 5- or 6-membered saturated heterocycles, such as 1-pyrrolidinyl, 4-morpholinyl, 1-piperidinyl, 1-piperazinyl or 1-pyrazolidinyl, 5- or 6-membered unsaturated heterocycles, such as 1-imidazolyl, and the like.

[0247] R 1 The alkyl, aryl, heterocyclic group may be optionally substituted by one or more substituents, wherein the substituents are the same or different and include lower alkyl; cycloalkyl, hydroxy; hydroxyC1-6 alkylene, such as hydroxymethyl, 2-hydroxyethyl or 3-hydroxypropyl; lower alkoxy; C1-6 alkoxyC1-6 alkyl, such as 2-methoxyethyl, 2-ethoxyethyl or 3-methoxypropyl; amino; alkylamino; dialkylamino; cyano; nitro; acyl; carboxyl; lower alkoxycarbonyl; halogen; mercapto; C1-6 alkylthio, such as methylthio, ethylthio, propylthio or Butylthio; substituted C1-6 alkylthio, such as methoxyethylthio, methylthioethylthio, hydroxyethylthio or chloroethylthio; aryl; substituted C6-10 monocyclic or condensed ring aryl, such as 4-hydroxyphenyl, 4-methoxyphenyl, 4-fluorophenyl, 4-chlorophenyl or 3,4-dichlorophenyl; 5-6 membered unsaturated heterocyclic ring, such as furanyl, pyrrolyl, pyrazolyl, imidazolyl, thiazolyl, thienyl, pyridyl or pyrimidinyl; and bicyclic unsaturated heterocyclic ring, such as indolyl, isoindolyl, quinolyl, benzothiazolyl, chromanyl, benzofuranyl or o-phthalimino. In certain embodiments, one or more of the above groups can be explicitly excluded as substituents for various other groups of the formula. In some embodiments, the five-membered ring of the formula is a thiazole ring.

[0248] R 2The alkyl, aryl, heterocyclic group may be optionally substituted by one or more substituents, wherein the substituents are the same or different and include hydroxyl; C1-6 alkoxy, such as methoxy, ethoxy or propoxy; carboxyl; C2-7 alkoxycarbonyl, such as methoxycarbonyl, ethoxycarbonyl or propoxycarbonyl) and halogen. c The alkyl, aryl, heterocyclic group may be optionally substituted by one or more substituents, wherein the substituents are the same or different and include C3-6 cycloalkyl; hydroxy; C1-6 alkoxy; amino; cyano; aryl; substituted aryl, such as 4-hydroxyphenyl, 4-methoxyphenyl, 4-chlorophenyl or 3,4-dichlorophenyl; nitro and halogen.

[0249] With R c and R 1 The heterocyclic ring formed by the C1-6 alkylene radical and the nitrogen atom to which they are attached may be optionally substituted by one or more substituents, wherein the substituents are the same or different and include C1-6 alkyl; hydroxy C1-6 alkylene; C1-6 alkoxy C1-6 alkylene; hydroxy; C1-6 alkoxy; and cyano.

[0250] In one embodiment, there is provided a micellar composition as defined herein, wherein: the TLR7 agonist has formula (IA); the amphiphilic micelle former is DSPE-PEG2000; and

[0251] The ratio between the TLR7 agonist and the amphiphilic micelle former was 95:5, 90:10 or 80:20.

[0252] In a preferred embodiment, there is provided a micellar composition as defined herein, wherein: the TLR7 agonist has formula (IA);

[0253] The amphiphilic micelle former is DSPE-PEG2000; and

[0254] The molar ratio between the amphiphilic micelle former and the TLR7 agonist was 95:5, 90:10 or 80:20.

[0255] Other active agents

[0256] In one embodiment, there is provided a micellar composition as defined herein, further comprising at least one additional active ingredient. In one embodiment, the micellar composition further comprises at least one antigen.

[0257] project

[0258] I-1. A micellar composition comprising:

[0259] A toll-like receptor 7 (TLR7) agonist of formula (I), formula (II), formula (III) or formula (IV);

[0260]

[0261] Where X 1 Is -O-, -S-, or –NR C ;

[0262] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0263] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or R C and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0264] R 2 each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl), halogen, nitro or cyano, or R 2 does not exist;

[0265] R a and R b each is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0266] Wherein the substituent on any alkyl, aryl or heterocyclic group is hydroxyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkylene, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 1-6 Alkoxy C 1-6 Alkylene, amino, cyano, halogen or aryl;

[0267] n is 0, 1, 2, 3, or 4;

[0268] X 2 is a bond or a linking group; and

[0269] R 3 It is a lipid;

[0270] X 3 is -N- or -CH-;

[0271] R 4 is -CH2- or -CH(R 2 )-;and

[0272] k is 0 or 1;

[0273] X 4 It is -O-, -S-, -NH-, -N(R d )-, -CH2- or -CH(R 2 )-;

[0274] R d each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -C(O)NR a R b (carbamoyl);

[0275] or a tautomer thereof;

[0276] or a pharmaceutically acceptable salt or solvate thereof, and

[0277] The ring system of formula (II) is a piperidine ring, one of the heteroatoms is a N atom and the N atom of the piperidine ring is 2 adjacent, and

[0278] wherein the purine group in any of Formulas (I), (II), (III) or (IV) undergoes tautomeric rearrangement;

[0279] and amphiphilic micelle formers.

[0280] I-2. A micellar composition according to any one of the preceding items, wherein the TLR7 agonist has formula (I):

[0281]

[0282] Where X 1 Is -O-, -S-, or –NR C ;

[0283] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0284] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or R C and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0285] R 2 each independently represents -OH, (C1-C6) alkyl, substituted (C1-C6) alkyl, (C1-C6) alkoxy, substituted (C1-C6) alkoxy, -C(O)-(C1-C6) alkyl (alkanoyl), substituted -C(O)-(C1-C6) alkyl, -C(O)-(C1-C6) alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl), halogen, nitro or cyano, or R 2 does not exist;

[0286] R a and R beach is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0287] Wherein the substituent on any alkyl, aryl or heterocyclic group is hydroxyl, C 1-6 Alkyl, hydroxyl C 1-6 Alkylene, C 1-6 Alkoxy, C 3-6 Cycloalkyl, C 1-6 Alkoxy C 1-6 Alkylene, amino, cyano, halogen or aryl;

[0288] n is 0, 1, or 2;

[0289] X 2 is a bond or a linking group; and

[0290] R 3 It is a lipid;

[0291] or a tautomer thereof;

[0292] or a pharmaceutically acceptable salt or solvate thereof, and

[0293] wherein the purine group undergoes tautomeric rearrangement.

[0294] I-3. A micellar composition according to any one of the preceding items, wherein the TLR7 agonist has formula (I):

[0295]

[0296] Where X 1 Is -O-, -S-, or –NR C ;

[0297] R 1 is hydrogen, (C1-C 10 ) alkyl, substituted (C1-C 10 ) alkyl, C 6-10 Aryl or substituted C 6-10 Aryl, C 5-9 Heterocyclic, substituted C 5-9 Heterocyclic ring;

[0298] R C It is hydrogen, C 1-10 Alkyl or substituted C 1-10 Alkyl; or RC and R 1 together with the nitrogen to which they are attached, form a heterocyclic or substituted heterocyclic ring;

[0299] R 2 does not exist;

[0300] n is 0;

[0301] X 2 is a bond or a linking group; and

[0302] R 3 It is a lipid;

[0303] or a tautomer thereof;

[0304] or a pharmaceutically acceptable salt or solvate thereof, and

[0305] wherein the purine group undergoes tautomeric rearrangement.

[0306] I-4. A micellar composition according to any one of the preceding items, wherein X 2 Selected from: a bond, -O-, -C(O)-(carbonyl), (C1-C6)alkyl, substituted (C1-C6)alkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, -C(O)-(C1-C6)alkyl (alkanoyl), substituted -C(O)-(C1-C6)alkyl, -C(O)-(C1-C6)alkyl 10 ) aryl (aroyl), substituted -C(O)-(C6-C 10 )aryl, -C(O)OH(carboxyl), -C(O)O(C1-C6)alkyl(alkoxycarbonyl), substituted -C(O)O(C1-C6)alkyl, -NR a R b 、-C(O)NR a R b (carbamoyl); and

[0307] R a and R b each is independently hydrogen, (C1-C6)alkyl, substituted (C1-C6)alkyl, (C3-C8)cycloalkyl, substituted (C3-C8)cycloalkyl, (C1-C6)alkoxy, substituted (C1-C6)alkoxy, (C1-C6)alkanoyl, substituted (C1-C6)alkanoyl, aryl, aryl(C1-C6)alkyl, Het, Het(C1-C6)alkyl, or (C1-C6)alkoxycarbonyl;

[0308] And among them

[0309] X 1 Is -O-, -S- or -NRC ;

[0310] R 1 is hydrogen, C1-C6 alkyl, or substituted (C1-C6) alkyl;

[0311] R C It is hydrogen, C 1-6 Alkyl or substituted C 1-6 Alkyl; or R C and R 1 Together with the nitrogen to which they are attached they form a heterocyclic or substituted heterocyclic ring.

[0312] I-5. A micellar composition according to any one of the preceding items, wherein R 3 It is a lipid selected from the group consisting of: phospholipids containing one or two carboxylic acid esters; steranes, such as cholesterol; glycolipids; and glycerides.

[0313] I-6. A micellar composition according to any one of the preceding items, wherein R 3 It is a phospholipid containing one or two carboxylic acid esters.

[0314] I-7. A micellar composition according to any one of the preceding items, wherein the diameter of the micelle is between 5nm and 50nm, such as between 6nm and 46nm, such as between 7nm and 42nm, such as between 8nm and 38nm, such as between 9nm and 34nm, such as between 10nm and 34nm, such as between 11nm and 30nm, such as between 12nm and 26nm.

[0315] I-8. A micellar composition according to any one of the preceding items, wherein the diameter of the micelle is between 5nm and 25nm, such as between 6nm and 24nm, such as between 7nm and 23nm, such as between 8nm and 22nm, such as between 9nm and 21nm, such as between 10nm and 20nm, such as between 11nm and 19nm, such as between 12nm and 18nm, such as between 13nm and 17nm, such as between 14nm and 16nm, such as 15nm.

[0316] I-9. The micellar composition according to any one of the preceding items, wherein the amphiphilic micelle former is selected from the group consisting of poloxamers, poloxamines, PEG-polyesters, PEG-polyanhydrides, PEG-polyamino acids, phospholipids, polysorbates and polyoxyethylene alkyl ethers.

[0317] I-10. The micellar composition according to any one of the preceding items, wherein the PEG-polyester is selected from: PEG-poly(lactic acid) (PEG-PLA), PEG-poly(lactic-co-glycolic acid) (PLGA) and PEG-poly(ε-caprolactone) (PCL).

[0318] I-11. The micellar composition according to any of the preceding items, wherein the PEG-polyanhydride is PEG-polysebacic anhydride (PSA).

[0319] I-12. A micellar composition according to any of the preceding items, wherein the PEG-polyamino acid is selected from: PEG-poly (L-histidine), PEG-poly (L-aspartic acid), PEG-poly (L-asparagine), PEG-poly (L-glutamic acid), PEG-poly (L-glutamine) and PEG-poly (L-lysine).

[0320] I-13. The micellar composition according to any one of the preceding items, wherein the amphiphilic micelle former is a phospholipid conjugated to polyethylene glycol (PEG).

[0321] I-14. The micellar composition according to any one of the preceding items, wherein the phospholipid conjugated to PEG is conjugated via a carbonyl group.

[0322] I-15. A micellar composition according to any one of the preceding items, wherein the size of the PEG is between PEG350 and PEG5000, for example, between PEG550 and PEG4000, for example, between PEG750 and PEG3000, such as between PEG1000 and PEG3000, preferably the size of the PEG is PEG2000.

[0323] I-16. A micellar composition according to any one of the preceding items, wherein the phospholipid comprises one or more alkyl chains, and the one or more alkyl chains are one or more C8-C24 alkyl chains, such as C10-C22, such as C12-C20, preferably C14-C18, most preferably C16-C18 saturated alkyl chains or unsaturated alkyl chains.

[0324] I-17. A micellar composition according to any one of the preceding items, wherein the phospholipid comprises phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidic acid (PA), diphosphatidylglycerol (DPG) or phosphatidyl alcohol.

[0325] I-18. A micellar composition according to any one of the preceding items, wherein the phosphatidylethanolamine is selected from 1,2-dioleoyl-phosphatidylethanolamine, 1,2-dipalmitoyl-phosphatidylethanolamine, 1,2-dimyristoyl-phosphatidylethanolamine, 1,2-distearoyl-phosphatidylethanolamine, 1-oleoyl-2-palmitoyl-phosphatidylethanolamine, 1-oleoyl-2-stearoyl-phosphatidylethanolamine, 1-palmitoyl-2-oleoyl-phosphatidylethanolamine and 1-stearoyl-2-oleoyl-phosphatidylethanolamine.

[0326] I-19. A micellar composition according to any one of the preceding items, wherein the phospholipid conjugated to PEG is selected from: 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)-PEG, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE)-PEG and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE)-PEG.

[0327] I-20. The micellar composition according to any one of the preceding items, wherein the phospholipid conjugated to PEG is: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG.

[0328] I-21. The micellar composition according to any one of the preceding items, wherein the amphiphilic micelle former is DSPE-PEG2000.

[0329] I-22. A micellar composition according to any one of the preceding items, wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 50:50 to 99.5:0.5, such as 60:40 to 99:1, such as 70:30 to 98:2, such as 80:20 to 95:5, for example 95:5, 90:10 or 80:20.

[0330] I-23. A micellar composition according to any of the preceding items, wherein the composition comprises a TLR7 agonist at a molar concentration between 1% and 25%, such as 1%, 2%, 3%, 4%, 5%, 6%, 7%, 8%, 9%, 10%, 11%, 12%, 13%, 14%, 15%, 16%, 17%, 18%, 19%, 20%, 21%, 22%, 23%, 24%, or 25%.

[0331] I-24. A micellar composition according to any one of the preceding items, wherein the TLR7 agonist has a structure of formula (IA):

[0332] or a tautomer thereof;

[0333] or a pharmaceutically acceptable salt or solvate thereof.

[0334] I-25. A micellar composition according to any one of the preceding items, wherein: the TLR7 agonist has formula (IA); the amphiphilic micelle former is DSPE-PEG2000; and the ratio between the TLR7 agonist and the amphiphilic micelle former is 95:5, 90:10 or 80:20.

[0335] I-26. The micellar composition according to any one of the preceding items, wherein the molar ratio between DSPE-PEG2000 and the TLR7 agonist of formula (IA) is 90:10 (MBS8).

[0336] I-27. The micellar composition according to any of the preceding items, further comprising at least one additional active ingredient.

[0337] I-28. The micellar composition according to any one of the preceding items, further comprising at least one antigen.

[0338] I-29. A pharmaceutical composition comprising the micellar composition according to any one of the preceding items.

[0339] I-30. The micellar composition or pharmaceutical composition according to any one of the preceding items, for use in preventing, treating or ameliorating a disease or disorder.

[0340] I-31. The micellar composition or pharmaceutical composition for use according to any of the preceding items, wherein the disease or disorder is selected from the group consisting of cancer, infectious diseases, inflammatory conditions or diseases, autoimmune diseases and allergies.

[0341] I-32. The micellar composition or pharmaceutical composition for use according to any one of the preceding items, wherein the disease or disorder is cancer, such as colon cancer.

[0342] I-33. A method for in vivo activation of immune cells of a subject, comprising administering to the subject the micellar composition or pharmaceutical composition according to any one of the preceding items in an amount sufficient to activate the immune cells.

[0343] I-34. A method for treating cancer in a patient in need of treatment, comprising administering to said patient a micellar composition as defined in any of the preceding items.

[0344] I-35. The method according to any of the preceding items, further comprising administering a chemotherapeutic agent to the patient.

[0345] I-36. A method according to any of the preceding items, wherein the chemotherapeutic agent is selected from doxorubicin, Doxil, epirubicin, cyclophosphamide, bortezomib and oxaliplatin.

[0346] I-37. A method according to any of the preceding items, further comprising administering an immune checkpoint inhibitor to the patient.

[0347] I-38. A method according to any of the preceding items, wherein the immune checkpoint inhibitor is selected from: atezolizumab, avelumab, durvalumab, nivolumab, tilelizumab, pembrolizumab and ipilimumab.

[0348] I-39. A method according to any of the preceding items, wherein the micellar composition is MBS8 and the immune checkpoint inhibitor is selected from: nivolumab and pembrolizumab.

[0349] I-40. The method according to any one of the preceding items, further comprising administering to the patient a monoclonal antibody targeting CD20.

[0350] I-41. The method according to any of the preceding items, comprising administering MBS8 and a monoclonal antibody targeting CD20.

[0351] I-42. The method according to any of the preceding items, further comprising administering to the patient a monoclonal antibody targeting epidermal growth factor receptor (EGFR).

[0352] I-43. The method according to any of the preceding items, comprising administering MBS8 and a monoclonal antibody targeting epidermal growth factor receptor (EGFR).

[0353] I-44. The method according to any one of the preceding items, further comprising administering to the patient a monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2).

[0354] I-45. The method according to any one of the preceding items, comprising administering MBS8 and a monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2).

[0355] I-46. The method according to any of the preceding items, further comprising administering to the patient a monoclonal antibody targeting CD38, such as daratumumab or isatuximab.

[0356] I-47. The method according to any of the preceding items, comprising administering MBS8 and a monoclonal antibody targeting CD38, such as daratumumab or isatuximab.

[0357] I-48. The method according to any of the preceding items, further comprising administering to the patient a monoclonal antibody targeting CD47, such as mololimab.

[0358] I-49. The method according to any of the preceding items, comprising administering MBS8 and a monoclonal antibody targeting CD47, such as mololimab.

[0359] I-50. The method according to any of the preceding items, further comprising radiotherapy.

[0360] Example

[0361] Example 1: Micelles and liposomes preparation

[0362] Micelles were prepared from 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000](DSPE-PEG2000) obtained from Lipoid GmbH. In brief, lipids were dissolved in tert-butyl alcohol: water (volume ratio of 9: 1) to a final concentration of 5-10 mM in a glass vial, and placed under magnetic stirring and heated to 50 degrees Celsius until completely dissolved. Solvents were removed by freezing vials in liquid nitrogen and then lyophilizing overnight. By dispersing the dried lipids in a buffer solution containing 150 mM NaCl, 10 mM phosphate (PH=7.4), the vials were exposed to slight vortexing to establish initial contact between lipids and solvents, and then exposed to ultrasonic treatment for 30 minutes to ensure that micelle structures were formed to prepare micelles. The dispersion was vortexed again, and then the dispersion was exposed to further ultrasonic treatment for 30 minutes. Micelles were stored at 4 degrees Celsius before use and / or characterization.

[0363] The monolayer fully hydrated liposomes were prepared from a mixture of 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine (POPC), 1-palmitoyl-2-oleoyl-sn-glycero-3-phosphoglycerol (POPG), 1,2-dioleoyl-3-trimethylammonium-propane (DOTAP), cholesterol (Chol), and 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DOPE-PEG2000) and 1v270 (C57H93N6O12P, Mw=1085.4, (2,3-di(oleoyl)-3-phosphoethanolamine)). The molar ratio of each lipid in the liposomes was MBS1:POPC:Chol:DOTAP:1V270:DOPE-mPEG2k (44.25:30:20:0.75:5), and MBS2:POPC:Chol:POPG:1V270:DSPE-PEG2k (44.25:30:20:0.75:5). All lipids were obtained from Avanti Polar lipids or Lipoid. Briefly, appropriate weights of POPC, POPG, Chol, DOTAP, 1V270 and DOPE-PEG2000 were dissolved in chloroform. The solvent was removed by a gentle N2 stream and the lipid film was dried overnight at low pressure to remove trace solvents. Multilamellar vesicles were prepared by dispersing the dried lipids in a buffer solution containing: 150 mM KCL, 10 mM HEPES (PH = 7.5), 1 mM NaN3, 30 μM CaCl2 and 10 μM EDTA. As described by Mayer et al., Biochim.Biophys.Acta, 858, 161-168, multilamellar vesicles were extruded 10 times through two stacked 100 nm pore polycarbonate filters.

[0364] Example 2: Preparation of micelles incorporating toll-like receptor 7 (TLR7) agonists

[0365] Micelles were made from 1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[methoxy(polyethylene glycol)-2000] (DSPE-PEG2000) obtained from Lipoid GmbH and the TLR7 agonist 1v270 (C57H93N6O12P, Mw=1085.4, (2,3-bis(oleoyloxy)propyl 2-(4-((6-amino-2-(2-methoxyethoxy)-8-oxo-7H-purin-9(8H)-yl)methyl)benzamido)ethyl phosphate). The chemical structure of 1v270 is summarized in Figure 2In brief, lipid is dissolved in tert-butyl alcohol: water (volume ratio is 9: 1), reaches final concentration in glass vial is 5-10mM (DSPE-PEG2000) or 1-3mM (1v270), and is placed under magnetic stirring and heated to 50 degrees Celsius until completely dissolved. Then two kinds of lipid dispersions are mixed to the desired ratio (95: 5 to 80: 20 DSPE-PEG2000: 1v270 molar ratio). Solvent is removed by freezing vials in liquid nitrogen, then lyophilizing overnight. By the dried lipid dispersion in the buffer solution containing 150mM NaCl, 10mM phosphate (pH=7.4), the vial is exposed to slight vortex to establish initial contact between lipid and solvent, then exposed to ultrasonic treatment for 30 minutes to ensure that micelle structure is formed to prepare micelle. Dispersion is vortexed again, and then dispersion is exposed to further ultrasonic treatment for 30 minutes. Micelles were stored at 4 degrees Celsius prior to use and / or characterization. Batch names, molar ratios, lipid composition, zeta potential (mV), size (nm) and polydispersity index (PDI) are summarized in Figure 1.

[0366] Example 3: Characterization of micelle size and surface charge depending on composition

[0367] Micelles as outlined in Figure 1 were prepared in an attempt to prepare a stable formulation of the TLR7 agonist 1v207 in an aqueous solvent, thereby allowing injection in saline buffer. Micelles were prepared as described in Example 1+2, and their size (diameter) was measured in nanometers (nm) by dynamic light scattering in a buffer consisting of 5% (w / w) glucose, 10mM HEPES, 1mM CaCl2, MilliQ water pH 7.4. Micelles without 1V207 had an average size of 11.5nm (based on particle distribution by number), while when the 1V270 content was increased from 5% to 20%, the micelles containing 1V270 had an average size in the range of 11.5nm to 13-14nm (based on particle distribution by number) ( Figure 1B and 1C The zeta potential of empty micelles was measured in glucose buffer with CaCl2 and was approximately -4 mV, but became more negative when the anionic 1V270 compound was incorporated into the micelles, thus decreasing to -8 mV for micelles with 10% 1V207 ( Figure 1C ). Size and zeta potential were measured on a Zetasizer from Malvern Instruments.

[0368] Example 4: Antitumor activity of micelles and liposomes containing 1V270 in the CT26 model

[0369] Tumor studies in the CT26 model (Adlard et al., Int J Cancer, 135, 820-829, 2014): Briefly, the CT26 model is a colon cancer model established as a subcutaneous model in Balb / C mice, which is frequently used as an immunocompetent tumor model for testing cancer drugs and immunotherapeutic drugs.

[0370] We used this model for the control group, the radiotherapy treatment group, starting on day 10 and administering 2 Gy per day for 5 days. Liposomes or micelles containing 1V270 were injected intravenously for five doses ( Figure 3A and 3B ). Liposomes containing 5% 1V270 were injected to achieve a total dose of 266 nmol 1V270 / mouse ( Figure 3A ). The micelles were injected at three doses of 50, 100 and 200 nmol 1V270 / mouse and showed significant tumor growth inhibition compared to radiotherapy alone. In addition, the lowest dose of 50 nmol showed complete remission in 2 / 10 mice, the 100 nmol dose showed complete remission in 4 / 9 mice, and the 200 nmol dose showed complete remission in 8 / 8 mice. In addition, almost all mice in complete remission were able to resist re-challenge with the same tumor cells 100 days later without tumor regrowth, demonstrating that the mice had developed an anti-tumor immune memory response. In monotherapy, MBS8 micelles were also very potent in showing anti-tumor activity ( Figure 3C ), in which doses of 50, 200 and 400 nmol of MBS8 were injected intravenously into mice on day 9 and injected 4 more times at 4-day intervals. The two highest doses showed tumor growth inhibition in all 10 mice in the group, which lasted until 2 weeks after the last dose (day 34). On day 41, three mice in the two high-dose groups had complete remission. When combined with anti-PD1 treatment, 50 nmol / mouse / dose of MBS8 micelles showed synergistic effects. This combination treatment showed significantly better anti-tumor activity than both anti-PD1 alone and MBS8 alone (p<0.05)( Figure 3D ). Anti-PD1 was administered IP 6 times on days 11, 15, 19, 23, and 27.

[0371] Example 5: Antitumor activity of micelles containing TLR7 agonists compared to liposomes in combination with radiation therapy in the CT26 model

[0372] Multiple tumor studies were performed on groups of mice treated with radiation therapy, as well as 7 groups of liposome-treated mice (40-266 nmol 1V270 injected per mouse) and 13 groups of micelle-treated mice (50-200 nmol 1V270 injected per mouse) in combination with radiation therapy. The number of mice in each group was 7-9, and the median tumor size was compared between the groups treated with radiation therapy alone and with radiation therapy and liposomes or micelles. The median survival time of mice treated with micelles or liposomes in each group is shown in Table 1. Figure 3E . The 4 micelle-treated groups showed a median survival of more than 100 days, indicating that more than half of the mice showed complete tumor remission (the observed median survival of mice showed complete tumor remission). This was not observed in any of the liposome + RT treated groups. Tumor growth (called T / C ratio) calculated when the liposome + RT and micelle + RT treated groups were compared with the group treated with RT alone and when the group treated with RT alone was sacrificed showed that the T / C ratio was below 50% in 10 of the 13 micelle groups, while only 1 of the 7 liposome-treated groups showed a T / C ratio below 50%. Using the Wilcoxon rank sum test, the differences were significant at the p<0.01 level ( Figure 3F The T / C value can be converted to % tumor growth inhibition by subtracting the T / C value from 100% ( Figure 3G ), where values ​​close to 100% indicate very strong tumor growth inhibition. Ten of the 13 micelle-treated groups showed tumor growth inhibition of more than 60%, while only one of the seven liposome-treated groups showed tumor growth inhibition of more than 50%.

[0373] in conclusion

[0374] In conclusion, micelles containing 1V270 showed significantly better tumor growth inhibition than the corresponding liposomes containing 1V270 (p<0.01, Wilcoxon rank sum test).

[0375] Example 6: Micelles show safer cytokine profiles and reduce the risk of cytokine release syndrome (CRS)

[0376] For all samples except PBS and vehicle, micelles MBS6, MBS7 and MBS8 containing the TLR7 agonist 1V270 were compared with empty micelles (MBS0 = vehicle), PBS and two different liposomes containing 1V270 by intravenous injection in Balb / C mice at a dose of 100 nmol 1V270. Plasma samples were collected 2 and 6 h after injection, and cytokines associated with anti-tumor immune responses and toxicity were measured by multiplex or ELISA analysis.

[0377] At 2 or 6 h after treatment, all three micellar formulations of MBS6-8 had significantly higher levels of interferon-γ ( Figure 4A ) and interferon α ( Figure 4B ) were all reduced, supporting the reduced risk of cytokine storm with the micellar formulation. IL-12p70, which is important for eliciting type 1 cytotoxic T cell responses, showed a slight but significant decrease for MBS6 and MBS8 treatments compared to MBS1 liposomes, but was still at levels of cytokines expected to elicit anti-tumor responses ( Figure 4C ). At the 2h time point, mice treated with MBS6-8 had reduced levels of IL-6 in the plasma, a key cytokine that triggers CRS, compared to liposomal MBS1 and MBS2. After 6h, all mice showed IL-6 levels that were lower than after 2h, and IL-6 levels were similar or slightly lower than liposomes, demonstrating that micelles showed an overall reduction in IL-6 levels, not just a delay. TNFα, which is also associated with the initiation of the CRS syndrome, was also measured, and in particular, micelles showed an overall cytokine reduction compared to MBS1, with MBS6 and MBS8 being the most significant. The cytokine IL-1b and chemokine GRO were measured, but did not show significantly different cytokine levels.

[0378] in conclusion

[0379] In conclusion, administration of micellar MBS6-8 in mice resulted in a safer cytokine profile and reduced risk of cytokine release syndrome (CRS) compared with the corresponding liposomal MBS1 and MBS2 containing 1V270.

[0380] Example 7: Micelles show better toxicological profile compared to liposomes containing 1V270

[0381] Multiple studies were conducted with liposomes of varying charge and 1V270 content ranging from 0.75%-5% 1V270 and at doses of 13-266 nmol / mouse / dose injected into each mouse in groups of 7-9 mice / group. In the first two injections on days 0 and 4, the mice tolerated the liposomes well without any signs of adverse events or toxicity. However, in the third injection on day 8, mice showed adverse events and toxicity in 81% of the studies conducted. Toxicity was associated with lack of movement, piloerection, weight loss, and poor overall health for a brief period of time starting 10-15 minutes after dosing and lasting 30-40 minutes. This observation was observed in 30 of 37 studies of liposomes across all dose ranges.

[0382] In contrast, micelles have not shown this toxicity in any of the studies, currently 30 out of 30 studies, indicating that micellar formulations of 1V270 show a better toxicity profile than liposomal formulations of 1V270.

[0383] in conclusion

[0384] In conclusion, administration of micellar MBS6-8 in mice resulted in a better toxicological profile compared to the corresponding liposomes containing 1V270.

[0385] Example 8: Micelles induce lower IgM and IgG anti-PEG antibodies when injected into mice

[0386] To explore the toxicity associated with liposomes containing 1V270 but not micelles containing 1V270, liposomes and micelles containing 1V270 were injected once into mice by intravenous injection, and blood samples were drawn over a 28-day period. Micelles without 1V270 induced low levels of anti-PEG IgM antibodies in mouse plasma when measured by an ELISA kit for anti-PEG IgM antibodies ( Figure 5A MBS8 micelles induced anti-PEG IgM molecules in plasma on day 5, but the level was less than half of the amount induced by liposomes containing 1V270 ( Figure 5A , MBS8 versus MBS liposomes). For anti-PEG IgG antibodies, which are generally more specific in induction and persist longer in mammals, induction was very high with liposomes 1-2 weeks after dosing and then declined but persisted throughout the study ( Figure 5B Compared with liposomes containing 1V270, MBS8 micelles induced much lower IgG anti-PEG antibodies ( Figure 5B , approximately 17-fold lower). This observation suggests that micelles induce lower levels of IgM and IgG anti-PEG responses and thus reduce immune-related toxicity in mice and potentially other mammals, which is important for reducing toxicity in patients treated with micelles and liposomes with immune-boosting agents.

[0387] Figure 5C The specificity of anti-PEG antibodies raised against MBS8 is shown.

[0388] Healthy mice were treated with a single dose of MBS8 micelles with a total dose of 200nmol 1V270. Blood was drawn on the 5th day after injection, and plasma was obtained. Then, the plasma was incubated with PBS, PEGylated liposomes, free mPEG2000 chains, DSPE-PEG micelles without 1V270, or MBS8 micelles. In all pre-incubations, the PEG concentration was 1μM. The plasma was then added to a microscope well containing fixed fluorescently labeled PEGylated liposomes and incubated for 10 minutes. The microscope wells were washed, and a fluorescent secondary antibody against mouse IgM was added. The liposomes were imaged with a confocal microscope to determine the surface density of IgM on a single liposome.

[0389] When plasma was pre-incubated with PBS (no competition), the surface density was approximately 60 A.U. When plasma was pre-incubated with free mPEG, DSPE-PEG micelles, or MBS8 micelles, similar IgM binding was measured and showed high IgM surface density (AU in the range of 60-70), indicating that there was no competition for binding to the anti-PEG IgM present in the plasma. When plasma was pre-incubated with PEG-liposomes (competition positive control), the binding of anti-PEG IgM to the immobilized PEG-liposomes was almost completely abolished, indicating strong competition for binding. Therefore, Figure 5A The anti-PEG IgM induced by MBS8 micelles detected by ELISA in the present study was only able to recognize PEG chains attached to the surface of a planar or liposome, but not PEG micelles (including MBS8) or free PEG chains.

[0390] in conclusion

[0391] Anti-PEG IgM generated against MBS8 micelles after injection into mice generally failed to recognize and bind to either MBS8 micelles or DSPE-PEG micelles, supporting that anti-PEG IgM antibodies potentially generated in patients are unlikely to bind to MBS8 after multiple injections.

[0392] Example 9: Treatment of mammals with cancer using immunostimulatory 1v270 micelles (non-antigen specific) In order to obtain immunostimulatory micelles suitable for cancer treatment, micelles can be generated by mixing 1v270: DOPE-PEG2k (10: 90 = MBS8). The compounds are mixed in an organic solvent and dried into a lipid film. The film is hydrated in a buffer suitable for intravenous administration, such as containing saline. The micelles are administered intravenously at intervals of, for example, 1-2 weeks to cancer patients with, for example, lung cancer, breast cancer, prostate cancer, HNC, leukemia, lymphoma or melanoma. Combinations with clinically approved treatments may enhance anti-tumor effects. In particular, in combination with immune checkpoint inhibitors, in combination with radiotherapy to enhance, for example, the abscopal effect of lung cancer patients, and in combination with mAb therapies such as rituximab and trastuzumab to enhance antibody-dependent cellular cytotoxicity (ADCC) to enhance the response to immunogenic cell death induced by certain chemotherapy such as doxorubicin, oxaliplatin, cyclophosphamide and mitoxantrone.

[0393] Example 10: Treatment of mammals with cancer using immunostimulatory 1v270 micelles (antigen specific)

[0394] To obtain an antigen-specific immune response, micelles are prepared as in Example 9, but with the addition of an antigenic peptide comprising all or part of the antigen of interest linked to a lipid anchor such as, for example, DOPE. The peptide antigen associated with the lipid anchor ensures adequate micelle association, as seen with a 25 amino acid peptide sequence from the MUC1 tumor antigen, where palmitoylated lysine residues ensure adequate liposomal association of the antigen (Sangha and Butts, Clin Cancer Res 2007; 13, 15 supp, 2007, 4652-54s). The antigen can be, for example, a MAGE antigen for the treatment of melanoma, PSA for the treatment of prostate cancer, a neoantigen, or a third antigen or combination of antigens. The antigen is administered together with 1v270 to a cancer patient in whom the loaded antigen is expected to be expressed in his tumor. The micelles are administered multiple times to the same patient to enhance the antigen-specific response, preferably at intervals of 1-2 weeks.

[0395] Example 11: Antitumor activity of the combination of MBS8 micelles and radiotherapy

[0396] Mice bearing CT26 subcutaneous tumors were treated with radiotherapy (RT) and micelles containing 50, 100 or 200 nmol 1V270. Starting from the 12th day after tumor inoculation, micelles were given intravenously every 4 days for a total of 5 treatments. Starting from the 12th day after tumor inoculation, 2Gy RT was given to the tumor-bearing flank every day for 5 consecutive days. The number of mice in each group was 8-10, and mice were re-challenged with CT26 on the opposite flank on day 101 after the initial inoculation. The data for the tumor growth curve are the mean tumor size ± SEM.

[0397] Combining 50 or 100 nmol MBS6 with RT provided only modest tumor control, whereas combining 200 nmol MBS6 with RT provided good tumor control ( Fig. 6A ). For RT alone, 1 / 9 mice were complete responders and 1 / 1 mouse rejected rechallenge. For 50 nmol MBS6 combined with RT, 2 / 9 mice were complete responders and 2 / 2 rejected rechallenge. For 200 nmol MBS6 combined with RT, 7 / 9 mice were complete responders and 7 / 9 mice rejected rechallenge ( Fig. 6A -B).

[0398] The combination of MBS7 and RT showed a synergistic effect. For 50 nmol MBS7 combined with RT, 5 / 9 mice were complete responders and 5 / 5 rejected rechallenge. For 100 nmol MBS7 as a monotherapy, 3 / 9 mice were complete responders and 3 / 3 rejected rechallenge. The combination of 100 nmol MBS7 with RT provided 8 / 10 complete responders and 3 / 3 mice rejected rechallenge, and provided significantly improved survival compared to monotherapy (p=0.02, Mantel-Cox test; Figure 6C -D).

[0399] The combination of 50 nmol MBS8 with RT provided 2 / 10 complete responders, and 2 / 2 mice rejected rechallenge. For 100 nmol MBS8 combined with RT, 4 / 9 mice were complete responders, and 4 / 4 rejected rechallenge. For 200 nmol MBS8 combined with RT, 8 / 8 mice were complete responders, and 7 / 8 rejected rechallenge. In addition, for the combination of MBS8 with RT, a dose dependency could be observed (p<0.3, Mantel-Cox test; Fig. 6E -F).

[0400] in conclusion

[0401] Micelles containing 1V270 at molar ratios of 80:20, 90:10, and 95:5 showed strong antitumor activity both in monotherapy and in combination with radiotherapy.

[0402] Example 12: Antitumor activity of MBS8 micelles in monotherapy

[0403] The efficacy of MBS8 micelles as monotherapy was investigated in the CT26 syngeneic subcutaneous colon cancer model. 3When the mean volume of 100 μg / mL was reached (day 0), randomization and treatment of mice began. Groups of 10 mice were treated with PBS as vehicle control or with 3 doses of MBS8 at 50, 200 or 400 nmol / mouse / injection by intravenous push ( Fig. 7A MBS8 was given q4d for a total of 5 injections starting from day 9 (days 9, 13, 17, 21, and 25). Tumor growth was measured twice a week. The average tumor volume of treated mice was Fig. 7A The tumor volumes of individual animals are shown in Figure 7B (1-4) Statistical analysis using the Wilcoxon rank sum test revealed that the efficacy of MBS8 at medium and high doses was significantly enhanced compared to the control (p<0.01). Eight out of 10 mice showed complete remission in the medium and high dose MBS8 treatment groups.

[0404] in conclusion

[0405] MBS8 micelles containing 10 molar amounts of 1V270 showed significant antitumor activity in the CT26 model at doses of 200 and 400 nmol and were highly potent in inducing complete remission.

[0406] Example 13: Antitumor activity of the combination of MBS8 micellar PD-1 therapy

[0407] The efficacy of the combination of MBS8 micelles and αPD-1 therapy was studied with IP injections of a fixed dose of 10 mg / kg of αPD-1 and escalating doses of 50, 200 and 400 nmol / mouse / injection of MBS8. The CT26 model and MBS8 treatment were performed as described in Example 12. MBS8 treatment started on day 9 and αPD-1 started on day 11 and then twice a week for three weeks ( Fig. 8A Low-dose MBS8 combined with αPD-1 showed significant tumor growth delay compared with low-dose MBS8 alone and αPD-1 alone (p<0.05) ( Fig. 8A and 8B , Figures 1-4). At the mid- and high-dose MBS8, the combination therapy initially showed no difference, as all 10 mice in each group showed either remission or very small tumors ( Fig. 8A However, on days 30-50, several tumors started to grow in the MBS8 monotherapy group, of which two tumors escaped ( Fig. 8A and 8B , Coordinate Figures 3-4). In the αPD-1 combination group, these escapers were not seen ( Figure 8B , Figures 7 and 8), and all tumors eventually disappeared or became very small (<30 mm3).

[0408] in conclusion

[0409] The combination of MBS8 micelles containing 1V270 and αPD-1 was highly potent, resulting in complete remission in at least 90% of treated mice bearing the CT26 model.

[0410] Example 14: Antitumor activity of the combination of MBS8 micelles and chemotherapy (doxorubicin and Doxil) in the CT26 model

[0411] The efficacy of MBS8 micelles in combination with standard of care chemotherapy, including doxorubicin and doxil, was investigated and compared with the efficacy of these chemotherapy drugs as monotherapy in the CT26 syngeneic subcutaneous colon cancer model. 3 When the mean volume of the mice reached 1.57 μg / kg (day 0), randomization and treatment of mice was started. Groups of 10 mice were each treated with PBS as vehicle control, doxorubicin (4 mg / kg), or a combination of MBS8 (400 nmol / mouse) and doxorubicin (4 mg / kg) ( Fig. 9A ), or doxil (4 mg / kg) or the combination of MBS8 (200 nmol / mouse) and doxil (4 mg / kg) ( Fig. 9C ) were treated with 40% paraformaldehyde (P < 0.05). All drugs were administered by intravenous push. Doxil or doxil was given q4d for 3 injections. MBS8 was given q4d for 5 injections. Doxil or doxil was given on days 10, 14, and 18; MBS8 was given on days 10, 20, 24, 28, and 32. Tumor growth was measured twice a week. The mean tumor volume of animals treated with doxorubicin and / or MBS8 is shown in Table 1. Fig. 9A The tumor volumes of individual animals are shown in Fig. 9B The mean tumor volumes of animals treated with doxil and / or MBS8 are shown in Fig. 9C The tumor volumes of individual animals are shown in Fig.9D Statistical analysis using the Wilcoxon rank sum test revealed that the efficacy of the chemotherapy drugs was significantly enhanced when combined with MBS8. All animals treated with the MBS8 / doxil combination showed complete responses, and 8 of the 10 complete responders were in the group treated with MBS8 / doxil; no complete responders were found in the single chemotherapy groups.

[0412] in conclusion

[0413] Micelles containing 1V270 at 10 molar levels significantly potentiated the efficacy of doxorubicin and doxil and resulted in complete responders.

[0414] Example 15: Antitumor activity of MBS8 micelles using intravenous administration and intratumoral administration

[0415] Mice bearing CT26 subcutaneous tumors were treated with radiation therapy (RT) and intravenous or intratumoral injections of MBS8. Starting from day 12 after tumor inoculation, MBS8 or lipid-matched micelle vehicles without the TLR7 agonist 1V270 were given every 4 days for a total of 5 treatments. Starting from day 12 after tumor inoculation, 2Gy RT was given to the tumor-bearing flank every day for 5 consecutive days. Due to volume limitations, intratumoral MBS8 injections were given at 100nmol 1V270, and intravenous MBS8 injections were given at 200nmol 1V270 ( Fig. 10A The number of mice per group was 8, and mice in complete remission were re-challenged with CT26 on the opposite flank on day 103 after the initial challenge. Data for tumor growth curves are mean tumor size ± SEM. Regardless of the route of administration, MBS8 provided good tumor control as a monotherapy and in combination with radiation therapy, but the intravenous route was slightly more potent than intratumoral administration ( Fig. 10A ). Intravenous injection of MBS8 alone resulted in 3 / 8 complete responders, of which 3 / 3 rejected only rechallenge. Intravenous injection of MBS8 in combination with RT resulted in 6 / 8 complete responders, of which 5 / 6 rejected only rechallenge. Intratumoral injection of MBS8 alone resulted in 3 / 8 complete responders, of which 3 / 3 rejected only rechallenge. Intratumoral injection of MBS8 in combination with radiation therapy resulted in 4 / 8 complete responders, of which 4 / 4 rejected only rechallenge.

[0416] In contrast, vehicle injection did not improve tumor control ( Fig. 10B ). Radiation therapy alone or in combination with vehicle did not result in any complete responders. Intratumoral administration ( Fig. 10C ) and intravenous administration group ( Fig. 10D ). For MBS8 treatment in monotherapy, the median survival days were 43.5 days for intratumoral administration and 63 days for intravenous administration. For the combination of MBS8 and radiation therapy, the median survival time was 56 days for intratumoral administration and 100+ days for intravenous administration, as 6 out of 8 mice had a complete remission.

[0417] in conclusion

[0418] MBS8 micelles containing 1V270 provided good tumor control when injected intravenously or intratumorally and showed synergy with radiation therapy regardless of the route of administration.

[0419] Example 16: Antitumor activity of the combination of MBS8 micelles and radiotherapy

[0420] Mice carrying MC38, EL4 or B16-F10 tumors were treated intravenously with radiotherapy (RT) and MBS8. 200nmol MBS8 or lipid-matched vehicles were given q4d, for a total of 5 treatments. For mice carrying EL4, treatment was started on the 7th day after inoculation, micelles were given q4d, and radiotherapy was given to the flank carrying tumors with 2Gy for 3 consecutive days, with 9-10 mice per group. For mice carrying MC38, treatment was started on the 10th day after inoculation, and radiotherapy was given to the flank carrying tumors with 2Gy for 5 consecutive days, with 9 mice per group. On the 80th day after the initial excitation, the complete responders carrying MC38 were re-excited on the relative flanks with MC38.

[0421] MBS8 provided no tumor control in EL4 as monotherapy, but combined with RT resulted in 4 / 10 complete responders. RT alone produced only 1 / 9 complete responders, and the combination of vehicle and RT produced 2 / 10 complete responders ( Fig.11A -B).

[0422] When combined with radiation therapy, MBS8 provided moderate tumor control in MC38-bearing mice. The combination of RT and MBS8 given q4d resulted in 3 / 9 complete responders, 2 / 3 of whom rejected rechallenge. Fig. 11C -D).

[0423] in conclusion

[0424] In conclusion, MBS8 provided good tumor control in EL4 and MC38 when combined with RT.

[0425] Example 17: Tolerance of MBS8 micelles in monkeys

[0426] The tolerance of MBS8 micelles was evaluated in cynomolgus monkeys (macaca fascicularis). MBS8 was administered to three naive male monkeys weighing about 3 kg, using the following dose escalation scheme every 14 days: 0.01 mg / kg → 0.03 mg / kg → 0.1 mg / kg → 0.3 mg / kg → 0.9 mg / kg → 2.7 mg / kg. The drug was administered by intravenous infusion of a total of 5 ml at a rate of 0.25 ml / min. During the adaptation period, blood samples were collected as baseline controls on the 14th day and the 7th day, and blood samples were collected on the 2nd, 4th, 8th, 24th, 72th and 14th days during the treatment period. Hematology was analyzed at 8h, 24h, 72h and 14th days. Blood chemistry was analyzed on the 14th day after each administration, i.e., before the next administration. C-reactive protein (CRP) was measured at 4h, 8h, 24h, 72h and 14th days after administration. Body temperature was monitored every day. Blood pressure was measured 2h, 4h, 8h after each administration and then daily until the next dose. Food consumption and body weight were checked daily - once a week.

[0427] All doses were well tolerated. A transient dose-dependent increase in CRP levels was detected 24 h after infusion, which returned to baseline levels within 3 days ( Fig. 12A There were no significant changes in body temperature throughout the study ( Fig. 12B ). Based on blood chemistry, there were no signs of hepatotoxicity or changes in electrolyte profile.

[0428] in conclusion

[0429] Intravenous infusion of MBS8 was well tolerated in cynomolgus monkeys over the dose range of 0.01-2.7 mg / kg.

[0430] Example 18: Antitumor activity of MBS8 micelles and aPD-1 in monotherapy and combination therapy in multiple syngeneic tumor models

[0431] The efficacy of MBS8 as monotherapy or in combination with an anti-PD1 monoclonal antibody (clone RMP1-14) was studied in a panel of 12 syngeneic mouse models ( FIG. 13 ).

[0432] Mice were inoculated subcutaneously with cancer cells; when the average tumor size reached about 100 mm 3Animals were randomly divided into 4 treatment groups with 10 animals in each group: Group 1, vehicle control (PBS); Group 2, anti-PD-1, 10 mg / kg; Group 3, MBS8, 300 micrograms / mouse; Group 4, combination of MBS8 and anti-PD-1. Vehicle or MBS8 was administered by slow intravenous push injection starting on the day of randomization on a q4d schedule for a total of 5 injections. Anti-PD-1 was administered intraperitoneally starting two days after randomization on a q4d schedule for a total of 6 injections. The results are summarized as follows Fig.13A TGI%, tumor growth inhibition, was calculated as: TGI (%) = 100 x (1-T / C), where T and C were the average tumor volumes of the treatment group and the control group on the day the control group was terminated.

[0433] Three distinct response patterns were observed:

[0434] (1) MBS8 is very potent in monotherapy, and thus the benefit of combination with anti-PD-1 cannot be noticed due to the excessively potent MBS8 activity ( Fig. 13B ). The tumors belonging to this category are: CT-26 (colon carcinoma), EMT-6 (breast cancer), A20 (B-cell lymphoma) and H22 (hepatoma).

[0435] (2) Anti-PD-1 and MBS8 both showed therapeutic activity in monotherapy. Combination therapy showed additive benefits ( Fig. 13C ). This was observed in Hepa1-6 (hepatoma) and M38 (colon cancer).

[0436] (3) Anti-PD-1 was inactive as a monotherapy, while MBS8 monotherapy showed efficacy. The combination of the two drugs produced synergistic activity by converting PD-1 non-responsive mice into responsive mice ( Fig.13D ). In this category are RM-1 (prostate cancer), Pan02 (pancreatic cancer), and Renca (kidney cancer).

[0437] For the EMT-6 model, complete responders from groups 3 and 4 that were tumor-free for at least 3 weeks were re-challenged with subcutaneous injections of EMT-6 cells on the contralateral flank. Mice were followed for 29 days. All re-challenged animals showed complete rejection of the re-challenged tumor ( Fig.13E In the CT26 model, mice that showed a complete response to MBS8 or the combination of MBS8 and anti-PD-1 therapy and rejected re-challenged tumors were analyzed for the presence of tumor-specific T cells ( Fig.13F, Elispot plot). Splenocytes from untreated mice bearing CT26 tumors, mice treated with MBS8 or MBS8 and anti-PD-1, or naive tumor-free mice were stimulated in vitro with AH-1 tumor-specific antigen peptides. IFNγ-producing cells were quantified using ELISPOT. All mice that rejected re-challenged tumors showed an increase in the number of IFNγ-positive cells in response to antigen stimulation, thus confirming the establishment of an immune memory response.

[0438] in conclusion : MBS8 showed therapeutic efficacy in monotherapy. In some tumors that were weakly responsive to anti-PD-1, combination treatment with MBS8 had an additive effect. In models that were unresponsive to anti-PD-1 treatment but responsive to MBS8, the latter sensitized tumors to anti-PD-1, and the drugs showed a strong synergistic effect. MBS8 led to the establishment of an immune memory response in monotherapy or in combination with anti-PD-1.

[0439] Example 19: Optimization of MBS8 dose regimen and correlation with anti-tumor activity in the CT26 model

[0440] The effect of dose schedule on the efficacy of MBS8 monotherapy was analyzed using the CT26 colon cancer syngeneic mouse model. Mice with established tumors were treated with 200 nmol / mouse MBS8 as (1) a single injection; (2) 2 injections q7d; (3) 3 injections q7d; (4) 4 injections q7d and (5) 5 injections q7d (Figure 14). As a control, vehicle (PBS) treatment and MBS8 treatment were used, administered as five injections q4d, a schedule that has shown good efficacy in previous studies.

[0441] A single injection has shown significant inhibition of tumor growth, while multiple weekly administrations resulted in complete tumor eradication.

[0442] in conclusion: Only a single injection of MBS8 resulted in significant antitumor activity, while additional injections over 4 or 7 days resulted in complete remissions in multiple mice.

[0443] Example 20: Antitumor activity of MBS8 micelles compared to R848 monotherapy

[0444] The efficacy of MBS8 micelles and R848 as monotherapy was investigated in the CT26 syngeneic subcutaneous colon cancer model. 3The randomization and treatment of mice was started when the mean volume of the tumor reached 1.5 μg / mL (day 11). Groups of 9 mice were each treated with vehicle control (MBS0); MBS8 at 100, 200 or 300 nmol / mouse / injection by intravenous push or R848 at 200 nmol / mouse / injection. Treatment was given q4d for a total of 5 injections starting on day 11 (days 11, 15, 19, 23 and 27). Tumor growth was measured twice a week. Fig.15A The mean tumor volume of treated mice is shown. Fig. 15B Survival curves are shown. Statistical analysis using the Wilcoxon rank sum test revealed significant differences for all doses of MBS8 compared to vehicle (p<0.0001). In addition, 200 nmol MBS8 was significantly more effective than 200 nmol R848 (p<0.05). Weight changes from baseline were as shown in Fig. 15C For MBS8 micelles, the weight loss changes became less severe with more treatments, while R848 induced the most severe weight loss changes from day 25 to day 40.

[0445] in conclusion

[0446] MBS8 micelles showed better antitumor effects than R848 when given at equimolar doses. Due to enhanced therapeutic activity and less severe weight loss, MBS8 micelles have an increased therapeutic index compared with R848, a TLR7 agonist.

[0447] Example 21: Analysis of tumor gene expression in MBS8-treated CT26 tumor-bearing mice using Nanostring

[0448] Gene expression profiles of CT26 tumors from mice treated with MBS8 as a monotherapy were made to determine the gene expression associated with MBS8 treatment and the presence of specific tumor cell types. Gene expression was evaluated using the Pan Cancer Immune Panel (Nanostring) performed on bulk RNA extracted from tumors. Mice were randomly divided into 10 groups on day 12 and treated with MBS8 at 200 nmol / mouse / injection by intravenous push, given on a q4d schedule, for 1 or 3 injections. Mice were sacrificed and tumors were snap frozen on the following days: day 0 (untreated), 1 day after the first injection, 2 days after the first injection, 4 days after the first injection, 14 days after the first injection, 2 days after the third injection, 4 days after the third injection, for all groups, n=3-6. RNA was extracted and 750 genes were analyzed using the Pan Cancer Immune Panel. Cell type analysis was performed using the Advanced Analysis Module of Nanostring software. Two-way ANOVA with multiple comparison correction was performed to compare different time points with untreated. At early time points (1, 2, and 4 days after the first injection), a significant increase in neutrophils and dendritic cells was observed, demonstrating activation of the innate immune system. At subsequent time points (4 days after the first injection and beyond), T cells and especially CD8 T cells in the tumor increased, demonstrating activation of the adaptive immune system. In addition, macrophages were also upregulated at later time points. The time point of 14 days after the first injection had a cell type profile similar to that of UT, indicating that a single injection in this setting was not sufficient to induce a lasting immune response.

[0449] in conclusion

[0450] Gene expression profiling of tumors from mice treated with MBS8 monotherapy showed induction of innate immune responses at early time points, with the presence of large numbers of neutrophils and dendritic cells. The adaptive immune response was dominated by T cells at later time points, which collectively mediated the antitumor effects of MBS8 treatment.

[0451] Example 22: Acute effects of MBS8 treatment on the tumor microenvironment

[0452] The acute effect of intravenous injection of 200 nmol MBS8 on tumors was evaluated by multicolor flow cytometry. Mice bearing CT26 tumors were treated with MBS8 and tumors were evaluated by flow cytometry at 1, 3 and 6 hours after injection. Red blood cells were determined based on size and Ter-119 expression and excluded from further analysis. All further analyses were determined based on the lack of Ter-119 expression and based on size and staining as live. CD8+T cells were defined as CD45+CD3+CD8+.

[0453] Six hours after MBS8 injection, red blood cells constituted the majority of cells in the tumor ( Fig.17A In addition, 6 hours after MBS8 injection, the tumor microenvironment was characterized by a reduction in immune cells (CD45+) and a significant and surprising decrease in cancer cells ( Fig. 17B , Figures 1 and 2). Despite the reduction of immune cells in the tumor, there was a strong increase in neutrophils 6 h after treatment ( Fig. 17C , Coordinate Figure 1), while T cells, NKT and NK cells did not change significantly ( Fig. 17C ,coordinate Figure 2 However, when immune activation of these cell subsets was monitored based on CD69 expression, T cells, NKT cells, and NK cells showed significant activation 3 h after treatment ( Fig.17D , coordinate diagram 1-5).

[0454] in conclusion

[0455] Intravenous injection of MBS8 induced an enrichment of neutrophils and erythrocytes in tumors 6 h after treatment, accompanied by a decrease in viable tumor cells and immune cells, while a significant activation of the following immune cells was seen: CD8 T cells, NKT cells, and NK cells.

[0456] Example 23: Effects of TLR7 micelles on tumor microenvironment and spleen

[0457] The effect of intravenous injection of MBS6 on the tumor microenvironment was evaluated by multicolor flow cytometry. Mice bearing CT26 tumors were treated with 200 nmol MBS6 every 4 days from day 15 after cancer cell inoculation, and tumors and spleens were analyzed two days after the second injection and 6 days after the first injection (day 21). Viable cells were identified based on cell size and lack of staining with viability dyes. All further analyses were based on viable cells. Neutrophils were identified as CD45 + CD11b + 6g + Tumor-associated macrophages (TAMs) were identified as CD45 + 6g - CD11c + CD11b + CD64 高 Patrolling monocytes (PMos) were identified as CD45 + 6g - CD11b + CD11c - Ly6c - CD64 + Monocytic myeloid-derived suppressor cells (Mo-MDSCs) were identified as CD45 +6g - CD11b + CD11c - Ly6c 高 Classical DC (cDC) 1 was identified as CD45 + 6g - CD11c + CD64 低 CD11b 低 XCR1 + cDC2 was identified as CD45 + 6g - CD11c + CD64 低 XCR1 - CD11b 高 Plasmacytoid DCs (pDCs) were identified as CD45 + 6g - CD11c + CD64 低 XCR1 - CD11b - Ly6c 高 Siglec-H + .CD8 + T cells were identified as CD45 + Side scatter ow CD3 + CD4 - CD8 + Calculate cells per 100 mg based on the population of interest, tumor weight, and total cells.

[0458] Compared with untreated tumors, tumor weights were lower ( Fig.18A , p=0.056). In addition, after treatment with MBS6, cell viability in tumors was significantly reduced, which was barely reflected in the spleen ( Fig.18B , Figures 1 and 2). In living cells, MBS6 induces immune cells (CD45 + ) strongly increased ( Fig.18C ). This is consistent with Fig. 17C This is the opposite of what is seen in , but can be explained by the fact that this time point is 6 days after the first treatment. Neutrophils are highly enriched in the tumor microenvironment, as can already be seen in Figure 17 (relative to the depletion of other cell types) after 6 h of MBS6 treatment, which is not reflected in the spleen ( Fig.18D , Figures 1 and 2). All other investigated immune populations and cancer cells were strongly reduced in the tumor microenvironment ( Fig.18E , coordinate diagram 1-8).

[0459] in conclusion

[0460] Intravenous injection of MBS6 strongly reduced cell viability in the tumor microenvironment, while not affecting viability in the spleen. In addition, MBS6 induced a strong increase in neutrophils in the tumor microenvironment, which was not observed in the spleen.

[0461] Example 24: Optimal dosing schedule for MBS8 and Doxil

[0462] To achieve optimal tumor control, the dosing schedule is critical when using a combination of MBS8 and chemotherapy. We wanted to test whether Doxil treatment combined with MBS8 improves, and in this case, how MBS8 should be dosed to achieve the best response ( Fig.19 Doxil treatment alone showed tumor growth delay but no mice had complete remission and had 80% tumor growth inhibition (TGI) compared to vehicle-treated mice ( Fig.19 ). When MBS8 administration was initiated two days after the last Doxil treatment (MBS8 post-dose), there was a significant tumor growth inhibition of 90% compared to vehicle-treated mice, and significant anti-tumor activity compared to Doxil alone (p=0.0007). However, when MBS8 administration was initiated on the same day as Doxil (MBS8 pre-dose + post-dose), all mice had a complete remission with a TGI of 98%, and significantly better anti-tumor activity compared to Doxil alone or MBS8 alone after Doxil treatment.

[0463] in conclusion

[0464] These data show that MBS8 immunotherapy is significantly more effective when administered prior to chemotherapy on the same day and then followed by treatment after chemotherapy has terminated, compared to when immunotherapy is initiated prior to chemotherapy.

[0465] Example 25: Micelles show no evidence of accelerated blood clearance (ABC)

[0466] In mice:

[0467] Figure 20 shows the mean (+ / -SD) plasma concentration-time profiles of MBS8 in female and male rats on day 1 (first dose) and day 15 (one week after the last dose of two doses separated by 1 week) after administration of 3 mg / kg / day MBS8 (1V270) by intravenous infusion over 1 hour on a linear scale (A) and a semi-logarithmic scale (B). This example demonstrates the indifferent kinetics of MBS8 administered after 1 day and 15 days (third dose), respectively. This observation supports that the micelles of the present invention do not induce accelerated blood clearance (ABC).

[0468] In cynomolgus monkeys:

[0469] Figure 21 shows the overlap of individual plasma concentration-time curves of 1V270 in 1 female and 1 male cynomolgus monkeys on day 1 (A, first dose) and day 15 (B, third dose after two weekly doses) after intravenous infusion administration of 0.3, 1 and 3 mg / kg / day MBS8 (1V270) within 1 hour on a linear scale. This example demonstrates the indifferent kinetics of MBS8 administered after the third dose after 1 day and 15 days, respectively. This observation supports that the micelles of the present invention do not induce accelerated blood clearance (ABC).

[0470] in conclusion

[0471] In both mice and cynomolgus monkeys, the micelles of the present disclosure did not induce accelerated blood clearance (ABC) when infused on day 1 (first dose) and day 15 (third dose). This supports the significantly improved therapeutic utility of the micelles, at least compared to liposomes known to induce ABC events.

Claims

1. A micellar composition comprising: A toll-like receptor 7 (TLR7) agonist having the structure of Formula (IA): or a tautomer thereof; or a pharmaceutically acceptable salt thereof, and an amphiphilic micelle former, the amphiphilic micelle former being a phospholipid conjugated to polyethylene glycol (PEG), and wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 50:50 to 99.5:0.5, and The diameter of the micelles is between 5 nm and 25 nm.

2. The micellar composition according to claim 1, wherein the phospholipid conjugated to PEG is conjugated via a carbonyl group.

3. The micellar composition according to claim 1, wherein the size of PEG is between PEG350 and PEG5000. The micellar composition according to claim 3 , wherein the size of PEG is PEG2000.

5. The micellar composition according to claim 1, wherein the phospholipid comprises one or more alkyl chains, and the one or more alkyl chains are one or more C8-C24 alkyl groups.

6. The micellar composition according to claim 1, wherein the phospholipid comprises one or more alkyl chains, and the one or more alkyl chains are C16-C18 saturated alkyl chains or unsaturated alkyl chains.

7. The micellar composition according to claim 1, wherein the phospholipid comprises phosphatidylethanolamine (PE), phosphatidylcholine (PC), phosphatidylserine (PS), phosphatidylglycerol (PG), phosphatidylinositol (PI), phosphatidic acid (PA), diphosphatidylglycerol (DPG) or phosphatidyl alcohol.

8. The micellar composition according to claim 7, wherein the phosphatidylethanolamine is selected from 1,2-dioleoyl-phosphatidylethanolamine, 1,2-dipalmitoyl-phosphatidylethanolamine, 1,2-dimyristoyl-phosphatidylethanolamine, 1,2-distearoyl-phosphatidylethanolamine, 1-oleoyl-2-palmitoyl-phosphatidylethanolamine, 1-oleoyl-2-stearoyl-phosphatidylethanolamine, 1-palmitoyl-2-oleoyl-phosphatidylethanolamine and 1-stearoyl-2-oleoyl-phosphatidylethanolamine.

9. The micellar composition according to claim 1, wherein the phospholipid conjugated to PEG is selected from: 1,2-dioleoyl-sn-glycero-3-phosphoethanolamine (DOPE)-PEG, 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG, 1,2-dimyristoyl-sn-glycero-3-phosphoethanolamine (DMPE)-PEG and 1,2-dipalmitoyl-sn-glycero-3-phosphoethanolamine (DPPE)-PEG.

10. The micellar composition according to claim 1, wherein the phospholipid conjugated with PEG is: 1,2-distearoyl-sn-glycero-3-phosphoethanolamine (DSPE)-PEG. The micelle composition according to claim 1 , wherein the amphiphilic micelle former is DSPE-PEG2000. 12 . The micelle composition according to claim 1 , wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 60:40 to 99:

1. 13 . The micelle composition according to claim 1 , wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 70:30 to 98:

2. 14 . The micelle composition according to claim 1 , wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 80:20 to 95:

5. 15 . The micellar composition according to claim 1 , wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 95:5, 90:10 or 80:

20. 16 . The micellar composition according to claim 1 , wherein the molar ratio between the amphiphilic micelle former and the TLR7 agonist is 90:

10.

17. The micellar composition of claim 1, wherein the composition comprises a TLR7 agonist at a molar concentration of between 1% and 25%.

18. A micellar composition comprising: A TLR7 agonist of formula (IA); or a tautomer thereof; or a pharmaceutically acceptable salt thereof; and DSPE-PEG2000; and The molar ratio between DSPE-PEG2000 and the TLR7 agonist was 95:5, 90:10 or 80:

20. 19 . The micellar composition according to claim 18 , wherein the molar ratio between DSPE-PEG2000 and the TLR7 agonist of formula (IA) is 90:

10.

20. The micelle composition according to claim 18, wherein the diameter of the micelle is 5 nm to 25 nm.

21. The micellar composition according to claim 18, wherein the TLR7 agonist is a TLR7 agonist of formula (IA):

22. The micellar composition according to any one of claims 1 to 21, further comprising at least one additional active ingredient.

23. The micellar composition according to any one of claims 1 to 21, further comprising at least one antigen.

24. A pharmaceutical composition comprising the micellar composition according to any one of claims 1 to 21.

25. Use of the micellar composition according to any one of claims 1 to 21 in the preparation of a medicament for preventing, treating or ameliorating cancer in a patient, wherein the cancer is liver cancer, pancreatic cancer, lymphoma, breast cancer, prostate cancer, kidney cancer and colon cancer.

26. The use according to claim 25, wherein the cancer is selected from liver cancer, pancreatic cancer, lymphoma, breast cancer and colon cancer.

27. The use according to claim 25, wherein the cancer is selected from prostate cancer and kidney cancer.

28. The use according to claim 25, wherein the cancer is colon cancer.

29. The use according to claim 25, wherein the treatment further comprises treatment with a chemotherapeutic agent.

30. The use according to claim 29, wherein the chemotherapeutic agent is selected from the group consisting of doxorubicin, Doxil, epirubicin, cyclophosphamide, bortezomib and oxaliplatin.

31. The use according to claim 25, wherein the treatment further comprises treatment with an immune checkpoint inhibitor.

32. The use according to claim 25, wherein the use further comprises treatment with α-PD-1, α-PD-L1 or α-CTLA-4.

33. The use according to claim 31, wherein the immune checkpoint inhibitor is selected from: atezolizumab, avelumab, durvalumab, nivolumab, tislelizumab, pembrolizumab and ipilimumab.

34. The use according to claim 31, wherein the micellar composition is the micellar composition according to claim 19 and the immune checkpoint inhibitor is selected from: nivolumab and pembrolizumab.

35. The use according to claim 25, wherein the treatment further comprises treatment with a monoclonal antibody targeting CD20.

36. The use according to claim 25, wherein the micellar composition is the micellar composition according to claim 19 and the treatment further comprises treatment with a monoclonal antibody targeting CD20.

37. The use according to claim 25, wherein the treatment further comprises treatment with a monoclonal antibody targeting epidermal growth factor receptor (EGFR).

38. The use according to claim 25, wherein the micellar composition is the micellar composition according to claim 19 and the treatment further comprises treatment with a monoclonal antibody targeting epidermal growth factor receptor (EGFR).

39. The use according to claim 25, wherein the treatment further comprises treatment with a monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2).

40. The use according to claim 25, wherein the micellar composition is the micellar composition according to claim 19 and the treatment further comprises treatment with a monoclonal antibody targeting human epidermal growth factor receptor 2 (HER2).

41. The use according to claim 25, wherein the treatment further comprises treatment with a monoclonal antibody targeting CD38.

42. The use according to claim 25, wherein the micellar composition is the micellar composition according to claim 19 and the treatment further comprises treatment with a monoclonal antibody targeting CD38.

43. The use according to claim 25, wherein the treatment further comprises treatment with a monoclonal antibody targeting CD47.

44. The use according to claim 25, wherein the micellar composition is the micellar composition according to claim 19 and the treatment further comprises treatment with a monoclonal antibody targeting CD47.

45. The use according to claim 25, wherein the treatment further comprises radiation therapy.

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