Use of iodized oil emulsion as radiosensitizer or lymph node tracer

Abstract

The present invention relates to the technical field of radioactive therapy, in particular to the use of an iodized oil emulsion as a radiosensitizer or a lymph node tracer. The iodized oil emulsion comprises an oil phase and an aqueous phase, wherein the oil phase comprises an iodized oil and has a volume percentage content in the iodized oil emulsion of 30-90%; and the volume percentage content of the iodized oil in the oil phase is 30% or above. In the present invention, it was unexpectedly discovered during the research process that after being prepared into an emulsion state, the iodized oil has an excellent radiosensitization effect. The iodized oil emulsion alone can serve as a radiosensitizer, and can effectively improve the effect of radiotherapy and increase the killing efficiency of X-rays with regard to tumor cells. The use of the iodized oil emulsion as a radiosensitizer provided in the present invention has important significance for the field of existing radiotherapy treatments.

Description

Application of iodized oil emulsion as a radiotherapy sensitizer or lymph node tracer

[0001] Cross-application

[0002] This application claims priority to Chinese Patent Application No. 202311297141X, filed on October 8, 2023, entitled “Application of iodized oil emulsion as a radiotherapy sensitizer or lymph node tracer,” the disclosure of which is incorporated herein by reference in its entirety. Technical Field

[0003] The present invention relates to the technical field of radiotherapy, in particular to the application of iodized oil emulsion as a radiotherapy sensitizer or lymph node tracer. Background Art

[0004] Radiation therapy is a treatment method that uses radiation or radioactive substances to treat cancer and other diseases. It can directly damage the DNA structure of cancer cells, thereby inhibiting their growth and division, achieving therapeutic effects. The treatment process is generally painless or minimal, allowing patients to lead a normal life during treatment. However, it still has disadvantages such as high radiation doses, significant side effects on healthy tissues, and, in particular, the strong resistance of tumor cells to radiation.

[0005] Currently, to address the strong radioresistance of tumor cells, radiosensitizers are often used to improve the effectiveness of radiotherapy. Commonly used radiosensitizers include irisin, sodium glycidazole, and NBTXR3, but these drugs all have significant drawbacks. For example, irisin and sodium glycidazole are administered intravenously or orally, which reduces their effectiveness. They also have significant toxic side effects and require continuous administration during radiotherapy, further increasing the harm to the patient. Furthermore, the hafnium oxide in NBTXR3 cannot be metabolized or degraded in the body, and the long-term toxicity of the heavy metal is unknown. Furthermore, NBTXR3 is relatively expensive.

[0006] Summary of the Invention

[0007] In order to solve the problems existing in the prior art, the present invention provides the use of iodized oil emulsion as a radiotherapy sensitizer or lymph node tracer.

[0008] Lipiodol is due to its excellent absorption for X-ray, and its clinical general use as CT contrast agent.Also have lipiodol to be used for tumor treatment process, generally use simultaneously with chemotherapeutics in the transcatheter arterial chemoembolization (TACE) of liver cancer, based on the characteristics of its natural target tumor, lipiodol mainly has the effect of being a drug carrier at this.The present invention unexpectedly found in the research process, although lipiodol itself does not possess the cell that kills tumor cell, does not possess the effect of radiotherapy sensitization yet, when it is prepared into emulsion form, use lipiodol emulsion separately and can play the effect of radiotherapy sensitization, can be used as radiotherapy sensitizer and use.This point exceeds the expectation of technical personnel fully.

[0009] The present invention has found that, compared to radiosensitizers such as hafnium oxide which is also based on high Z elements, iodine has good clinical safety and metabolizability. Using lipiodol emulsion as a radiosensitizer can ensure clinical safety application and improve radiotherapy effect. Specifically, after adding lipiodol emulsion in radiotherapy, using the same dose of X-rays can cause stronger cell damage, reflecting the enhancement effect. After entering the body, lipiodol emulsion can be retained in, for example, liver cancer lesions for a long time, but can be quickly removed in normal liver tissue or cirrhosis of the liver tissue, therefore has a very safe application prospect. And in the lesion, lipiodol emulsion can be evenly targeted to the tumor site, and stays for a long time (for example, more than 1 month), realizing the needs of clinical single or multiple radiotherapy, continuous or every other day radiotherapy.

[0010] The lipiodol emulsion, a radiosensitizer provided by the present invention, can be used in conjunction with various cancer treatment options. Examples include surgery, interventional therapy, chemotherapy, or treatments involving cytostatics, cytotoxic drugs, targeted therapies, immunotherapy, vaccines, radionuclides (particularly immunoradionuclide), and any other biological or inorganic products designed to treat cancer. Furthermore, it can target lymph nodes and lymphatic vessels, serving as a lymph node and lymphatic vessel tracer to achieve radiotherapy clearance of lymph node and lymphatic vessel tumors.

[0011] In a first aspect, the present invention provides a radiotherapy sensitizer, which is composed of an oil phase and an aqueous phase; the oil phase includes iodized oil, and the volume percentage of the oil phase in the iodized oil emulsion is: 30-90%; the volume percentage of the iodized oil in the oil phase is above 30%.

[0012] Furthermore, the volume percentage of the oil phase in the iodized oil emulsion is 50-85%, preferably 70-85%.

[0013] Furthermore, the volume percentage of iodized oil in the oil phase is above 50%.

[0014] Furthermore, the radiotherapy sensitizer consists of an oil phase, an aqueous phase and a surfactant; the oil phase includes iodized oil, and the volume percentage of the oil phase in the iodized oil emulsion is 30-90%.

[0015] The iodized oil of the present invention can be one or more of iodized oil, iodized oil soft capsule or iodized vegetable oil in iodized oil injection, and can be purchased commercially.

[0016] The iodized oil emulsion of the present invention can be a water-in-oil emulsion, an oil-in-water emulsion, a water-in-oil-in-water emulsion or an oil-in-water-in-oil emulsion.

[0017] The particle size of the dispersed phase of the iodized oil emulsion of the present invention is 1nm-50μm.

[0018] Furthermore, the oil phase further comprises: one or more of vegetable oil, glycerol, octadecyl alcohol or stearic acid; and / or the aqueous phase comprises: one or more of non-ionic contrast agent solution, sodium chloride solution, water, glucose aqueous solution, Ringer's solution, sodium lactate Ringer's solution, acetate Ringer's solution, compound sodium acetate Ringer's solution, sodium bicarbonate Ringer's solution, hydroxyethyl starch, succinylated gelatin, dextran or sugar saline.

[0019] The reagent used in the aqueous phase of the present invention can be a commonly used clinical liquid, which not only facilitates the preparation of the iodized oil emulsion, reduces the production cost of the iodized oil emulsion, but also makes the iodized oil emulsion suitable for clinical requirements.

[0020] Furthermore, the non-ionic contrast agent solution includes one or more of iohexol, iopromide or ioversol.

[0021] Furthermore, the surfactant includes:

[0022] One or more of nonionic surfactants, anionic surfactants, cationic surfactants, natural surfactants, liposomes and polymers.

[0023] Furthermore, the nonionic surfactant includes Tween and / or polyethylene glycol; the anionic surfactant includes sodium dioctyl succinate sulfonate and / or sodium lauryl sulfate; the natural surfactant includes one or more of gum arabic, gelatin, or shellac; and the polymer includes one or more of polyethylene glycol, chitosan, polylactic acid-co-glycolic acid copolymer, hydroxypropyl methylcellulose, polyvinyl pyrrolidone, polymethyl methacrylate, or hydroxypropyl methylcellulose acetate succinate. Macromolecules such as chitosan can help form Pickering emulsions, increasing their dispersion and stability.

[0024] In a second aspect, the present invention provides a radiotherapy drug preparation, comprising:

[0025] A drug or a prodrug thereof having a chemotherapy effect, and the radiotherapy sensitizer.

[0026] The radiotherapy sensitizer provided by the present invention can carry drugs with chemotherapy effects. If the drug is hydrophilic, it can be dissolved in the aqueous phase; if the drug is hydrophobic, it can be dispersed in iodized oil.

[0027] Furthermore, the drug having a chemotherapy effect includes any one or more of the following:

[0028] i) Compounds that affect tissue growth, cell growth or cell differentiation;

[0029] ii) compounds that induce biological effects;

[0030] iii) Nanomedicines with the ability to promote radiosensitization or induce cellular changes.

[0031] The biological effects described in the present invention are biological effects that are beneficial to the effect of chemotherapy, such as immune response.

[0032] Nano medicine of the present invention can be granular nano material, and granular nano material can be used as surfactant to make pickering emulsion, also can be used as medicine performance therapeutic effect.Granular nano material of the present invention can be hydrophilic or hydrophobic nano particle (particle diameter is 1-1000nm).Can mix lipiodol emulsion and granular nano material by ultrasonic method, obtain the lipiodol emulsion that comprises granular nano material.

[0033] In a preferred embodiment of the present invention, the drug included in the lipiodol emulsion may include one or more of an antimicrobial agent and an antitumor agent. Furthermore, the antimicrobial agent may be an antibiotic, an antiviral agent, or an antifungal agent; the antitumor agent may be an anticancer chemotherapeutic agent (e.g., a cytostatic agent, a cytotoxic agent), as well as any other biological or inorganic product used to treat cancer, such as therapeutic nucleic acids, particularly microRNA (miRNA), short hairpin RNA (shRNA), and small interfering RNA (siRNA).

[0034] In a preferred embodiment of the present invention, the drug in the iodized oil emulsion includes one or more of a chemotherapeutic drug, a small molecule drug, a targeted drug, a protein / antibody drug, a photosensitizer or an immune-related drug.

[0035] In a preferred embodiment of the present invention, the drug in the iodized oil emulsion can be one or more of an anti-tumor vascular drug, an anti-tumor targeted drug or an anti-tumor nano drug.

[0036] Further, chemotherapy drugs include nimustine, carmustine, lomustine, cyclophosphamide, ifosfamide, glyphosate mustard semustine, ilisimol, doxorubicin, fluorouracil, mercaptopurine, thioguanine, cytarabine, azacitidine, paris saponin VI, fluguanidine, tegafur, gemcitabine, carmofur, hydroxyurea, methotrexate, eufortin, ancitabine, decitabine, actinomycin D, doxorubicin, daunorubicin, epirubicin, mitomycin, peclocycline, bleomycin, salinomycin, pirarubicin, irinotecan, Harringtonine, hydroxycamptothecin, vinorelbine, paclitaxel, taxotere, topotecan, vincristine, vindesine, vinblastine, teniposide, etoposide, atamestane, aminoglutethimide, letrozole, formestane, medroxyprogesterone acetate, tamoxifen, asparaginase, carboplatin, cisplatin, miplatin, dacarbazine, oxaliplatin, mitoxantrone, thioridazine, tirapazamine, motesafin gadolinium, procarbazine, boron carrier (small molecule boron drug of boron-10 or biological complex combined with boron) and one or more of the derivatives of the above drugs.

[0037] Furthermore, protein drugs include pembrolizumab, necituzumab, nivolumab, pembrolizumab, atezolizumab, durvalumab, ramucirumab, bevacizumab, lung cancer vaccine - CimaVax, Adotrastuzumab, anti-PD-L1, anti-CTLA-4, TNFerade, HSV-tk, G207, pertuzumab, trastuzumab, cetuximab, panitumumab, ipilimumab, emtansine One or more of ine, Lanreotideacetate, avelumab, Lu177dotatate, 25Sipueucel-T, recombinant human interleukin-2, atezolizumab, obinutuzumab, ofatumumab, rituximab, blinatumomab, ibritumomab tiuxetan, brentuximab daratumumab, cetuximab, elotuzumab, olaratumab, and recombinant human endostatin.

[0038] Furthermore, the anti-tumor vascular drugs include one or more of tyrosine kinase inhibitors, sorafenib, apatinib, axitinib, sunitinib, lenvatinib, regorafenib, pazopanib, vandetanib, nintedanib, cabozantinib, cediranib, anlotinib, fruquintinib, tivozanib, mosatinib and derivatives of the above drugs.

[0039] Furthermore, targeted anti-tumor drugs include gefitinib, erlotinib, osimertinib, afatinib, ceritinib, Alunbrig, brigatinib, alectinib, crizotinib, trametinib, dalatinib, palbociclib, ribociclib, pomalimab, neratinib, lapatinib, everolimus, veliparib, olaparib, salvicin, aflibercept, imatinib, sunitinib, niraparib, everolimus, temsirolimus, ponatinib, dasatinib, bortezomib, niraparib, temsirolimus, niraparib ... One or more of Sutinib, Midostaurin, Ibrutinib, Idelalisib, Venetoclax, NU-7441, KU-55933, EZN-2968, CBL0137HCl, Belinostat, Panobinostat, Romidepsin, Vorinostat, Bortezomib, Carfismitrol, Ixazomib, Panostatin, Denosumab, Cobimetinib, Alitretinoin, Vismodur, Sonidegi, and derivatives thereof.

[0040] The radiosensitizer described herein can be administered locally via various interventional routes, particularly transdermal administration, puncture, endoscopic administration, direct tissue injection, intramuscular injection, lymphatic and lymphatic vessel injection, and cervical canal administration. Local administration allows the radiosensitizer to be targeted and diffused to the tumor site, achieving long-term retention, providing a foundation for long-term, multiple-dose radiotherapy in clinical practice.

[0041] The radiotherapy sensitizer of the present invention can be prepared by one or more of the following methods: medical three-way drug mixing method, ultrasonic method, microfluidic method, membrane high-pressure extrusion method, microfluidic high-pressure reaction method, high-pressure homogenization method, supercritical fluid high-pressure method, high-pressure electrostatic spray method, or subcritical fluid high-pressure method to achieve emulsification of lipiodol. Furthermore, if drug loading is required, the drug can be dissolved or uniformly dispersed in the lipiodol emulsion under high pressure; the drug can also be combined with the lipiodol by a chemical reaction method with a stable chemical bond or connected through an intermediate connector.

[0042] In a third aspect, the present invention provides the use of iodized oil emulsion as a radiotherapy sensitizer or lymph node tracer (also including the use in the preparation of a radiotherapy sensitizer or lymph node tracer);

[0043] The iodized oil emulsion comprises: an oil phase and an aqueous phase;

[0044] The oil phase comprises iodized oil, and the volume percentage of the oil phase in the iodized oil emulsion is 30-90%; the volume percentage of the iodized oil in the oil phase is above 30%.

[0045] Furthermore, the oil phase further comprises: one or more of vegetable oil, glycerol, octadecyl alcohol or stearic acid; and / or the aqueous phase comprises: one or more of: non-ionic contrast agent solution, sodium chloride solution, water, glucose aqueous solution, Ringer's solution, sodium lactate Ringer's solution, acetate Ringer's solution, compound sodium acetate Ringer's solution, sodium bicarbonate Ringer's solution, hydroxyethyl starch, succinylated gelatin, dextran, sugar saline, etc.

[0046] Furthermore, the iodized oil emulsion also includes a surfactant.

[0047] Furthermore, the surfactant includes:

[0048] One or more of nonionic surfactants, anionic surfactants, cationic surfactants, natural surfactants, liposomes and polymers.

[0049] The radiotherapy described herein is radiation therapy, wherein radiation refers to ionizing radiation, including radiation caused by X-rays, alpha rays, beta rays, gamma rays, electron beams, neutron beams, ion beams, radioisotopes, and the like. X-rays are a particularly preferred excitation source; the radiation voltage is typically about 2 keV to 25,000 keV, further about 2 keV to 6,000 keV (linear accelerator sources), and even further about 2 keV to 1,500 keV (e.g., cobalt-60 sources). The X-ray radiation dose can be 1-80 Gy.

[0050] The radiotherapy of the present invention is applicable to tumor radiotherapy of organisms, especially mammals, which may be humans or mice.

[0051] The radiotherapy described herein is suitable for the radiotherapy of various tumor types, exhibiting a favorable radiosensitization effect on various tumors, resulting in a significant reduction in tumor size. These tumors may include one or more of skin malignancies (skin cancer), central nervous system tumors, head and neck tumors, lung cancer, breast cancer, gastrointestinal tumors, male genitourinary tumors, gynecological tumors, adrenal and retroperitoneal tumors, bone and soft tissue sarcomas (regardless of location), lymphomas, myelomas, leukemias, and childhood tumors.

[0052] Furthermore, skin cancer includes AIDS-related malignancies and melanoma; central nervous system tumors include tumors related to the brain, brainstem, cerebellum, pituitary gland, spinal canal, eyes, and orbits; gastrointestinal tumors include liver cancer and hepatobiliary tract cancer, colon cancer, rectal cancer and anal cancer, stomach cancer, pancreatic cancer, and esophageal cancer; male genitourinary system tumors include prostate cancer, testicular cancer, penile cancer, and urethral cancer; gynecological tumors include cervical cancer, endometrial cancer, ovarian cancer, fallopian tube cancer, vaginal cancer, and vulvar cancer; and childhood tumors include Wilm's tumor, neuroblastoma, central nervous system tumors, or Ewing's sarcoma.

[0053] The radiotherapy described in the present invention is suitable for the treatment of primary tumors or secondary invasions, local or distant metastases, as well as in the context of prevention to avoid secondary malignant central nervous system damage, such as invasion (metastasis) from melanoma, lung cancer, kidney cancer, breast cancer, etc.

[0054] Furthermore, the radiosensitizer can be used at any time during radiotherapy, for example, as a neoadjuvant (before cancer surgery) or as an adjuvant (after surgery), and can also be used for advanced tumors that cannot be removed by surgery.

[0055] The lipiodol emulsion of the present invention has high radiation absorption, high tumor targeting, long tumor diffusion and retention time, and good clinical safety. Using the lipiodol emulsion of one embodiment of the present invention as a radiotherapy sensitizer in conjunction with radiotherapy can reduce tumor size and prolong the life span of animals.

[0056] The present invention has the following beneficial effects:

[0057] During research, the present invention unexpectedly discovered that although iodized oil itself does not have a radiosensitizing effect, it can achieve excellent radiosensitization effects when prepared as an emulsion. Based on this, the present invention provides the use of iodized oil emulsion as a radiosensitizer.

[0058] The application of the iodized oil emulsion provided by the present invention has high radiation absorption, high tumor targeting, high tumor diffusion and retention time, and good clinical safety. Its effect of reducing tumor volume and prolonging the life cycle of animals is even better than that of existing hafnium oxide radiotherapy sensitizers, and its safety factor is much higher than that of hafnium oxide, which is of great significance in the field of radiotherapy. BRIEF DESCRIPTION OF THE DRAWINGS

[0059] In order to more clearly illustrate the technical solutions in the present invention or the prior art, a brief introduction is given below to the drawings required for use in the embodiments or the description of the prior art. Obviously, the drawings described below are some embodiments of the present invention. For ordinary technicians in this field, other drawings can be obtained based on these drawings without paying any creative work.

[0060] FIG1 is a schematic diagram showing a comparison of the X-ray absorption capabilities of hafnium oxide (HfO 2 ), iodized oil (IO), IG 2:1, and IG 4:1, provided in Example 2 of the present invention.

[0061] Figure 2 is a schematic diagram comparing the DLS test results of iodized oil, IW2:1, IW3:1, IW4:1, IG2:1, IG3:1, IG4:1 and IOE provided in Example 2 of the present invention; wherein a is the DLS test result of iodized oil, IW2:1, IW3:1, IW4:1, IG2:1, IG3:1 and IG4:1, and b is the DLS test result of IOE.

[0062] 3 is a schematic diagram showing the radiosensitization effect of each group on liver cancer provided by Example 2 of the present invention, including a blank control, X-rays alone, and X-rays combined with iodized oil (IO), hafnium oxide (HfO2), IG2:1, IW4:1, or IG4:1.

[0063] FIG4 is a schematic diagram showing the sensitization effect of radiotherapy on breast cancer in each group, including blank control, X-ray alone, and X-ray combined with iodized oil (IO), IG2:1, IW4:1, or IG4:1, provided in Example 2 of the present invention.

[0064] 5 is a schematic diagram showing the sensitization effect of iodized oil emulsion IOG on radiotherapy of pancreatic cancer provided in Example 2 of the present invention.

[0065] 6 is a schematic diagram of the sensitization effect of iodized oil emulsion IOG on radiotherapy of breast cancer provided in Example 2 of the present invention.

[0066] Figure 7 is a schematic diagram of the radiosensitization effects of different treatments on mouse tumors provided in Example 2 of the present invention; wherein a is the radiosensitization effect of IW2:1 loaded cisplatin drug on mouse tumors; b is a blank control (Ctrl in the figure), drug-loaded iodized oil emulsion (IOD in the figure), X-ray only (X-ray in the figure), pure drug (gemcitabine saline solution, D) + X-ray (D+X in the figure), iodized oil emulsion (IOE) + X-ray (IOE+X in the figure), and drug-loaded iodized oil emulsion (IOD) + X-ray (IOD+X in the figure); C is the full scan section results of tumors in the D+X group and the IOD group.

[0067] FIG8 is a schematic diagram showing the radiosensitization effect of iodized oil on different types of cancer cells provided in Example 2 of the present invention.

[0068] FIG9 is a schematic diagram of IO, IW4:1 and IG4:1 targeting and retention in lymph nodes provided in Example 2 of the present invention.

[0069] 10 is a schematic diagram of the radiosensitization effect of iodized oil emulsion IOG on lymph node size provided in Example 2 of the present invention.

[0070] FIG11 is a schematic diagram comparing the intratumoral distribution of the hafnium oxide group and the IG4:1 group provided in Example 2 of the present invention.

[0071] 12 is a schematic diagram of the radiotherapy effects of the blank control, iodized oil (IO), X-ray group, IO + X-ray group, and iohexol + X-ray group provided in Example 2 of the present invention.

[0072] FIG13 is a diagram showing the imaging results of mouse lung sections of the blank control, iodized oil (IO) and iodized oil emulsion (IOE) provided in Example 2 of the present invention. DETAILED DESCRIPTION

[0073] To make the objectives, technical solutions, and advantages of the present invention more clear, the technical solutions of the present invention will be clearly and completely described below in conjunction with the accompanying drawings. Obviously, the embodiments described are only some of the embodiments of the present invention, not all of them. Based on the embodiments of the present invention, all other embodiments obtained by ordinary technicians in this field without making creative efforts shall fall within the scope of protection of the present invention.

[0074] Example 1

[0075] This embodiment provides different types of iodized oil emulsions, as follows:

[0076] 1. Iodized oil and iopromide solution were mixed by a three-way method at a volume ratio of 2:1, 3:1 or 4:1 to prepare emulsions, which were named IW2:1, IW3:1 and IW4:1 respectively.

[0077] The iodized oil used in this embodiment was commercially available. Figures 1-4, 6, 9, and 11 are of the Hengrui brand, and the iodized oil used in other figures is produced by Guerbet, France (the iodine concentration in the currently available iodized oil products is about 480 mg / ml).

[0078] 2. Iodized oil was mixed with 0.9% sodium chloride or 5% glucose solution using a three-way method at different volumes. The emulsion obtained by mixing iodized oil and 5% glucose solution was abbreviated as IG. Specifically, iodized oil was mixed with 5% glucose solution using a three-way method at volume ratios of 2:1, 3:1, and 4:1 to prepare emulsions, which were named IG2:1, IG3:1, and IG4:1, respectively.

[0079] 3. Add surfactant to prepare iodized oil emulsion, which is as follows (% are all volume percentages):

[0080] (1) 80% oil phase (40% iodized oil + 60% soybean oil) + 20% water for injection + 5% emulsifier polyoxyethylene hydrogenated castor oil HCO-40;

[0081] (2) 60% iodized oil + 40% 5% glucose aqueous solution + 5% emulsifier (20% Tween 80 + 80% polyoxyethylene hydrogenated castor oil HCO-40);

[0082] (3) 50% iodized oil + 50% water for injection + 2% surfactant (20% Tween 80 + 80% polyoxyethylene hydrogenated castor oil HCO-40) + 1% chitosan.

[0083] Each emulsion was ultrasonically disrupted for 20 min and homogenized for 10 min.

[0084] 4. Add surfactant and chemotherapy drugs to prepare drug-loaded iodized oil emulsion, as follows (% are all volume percentages):

[0085] Gemcitabine hydrochloride (McLean) was first sonicated in normal saline. An emulsion was then prepared using a ratio of 60% lipiodol + 40% normal saline + 5% emulsifier (20% Tween 80 + 80% polyoxyethylene hydrogenated castor oil HCO-40). The emulsion was sonicated for 20 minutes and homogenized for 10 minutes. The undosed lipiodol emulsion was labeled IOE, while the drug-loaded lipiodol emulsion (drug concentration 35 mg / mL) was labeled IOD.

[0086] Example 2

[0087] In this example, the iodized oil emulsion involved in Example 1 was subjected to X-ray attenuation coefficient comparison and radiotherapy sensitization performance test, as follows:

[0088] 1. The X-ray absorption capacity of hafnium oxide (HfO2), iodized oil (IO), IG2:1 and IG4:1 was tested through CT value comparison test.

[0089] Among them, hafnium oxide (HfO2) is synthesized according to the method provided by NANOBIOTIX company patent (patent number WO2009147214).

[0090] The results are shown in Figure 1. The CT values ​​of iodized oil, IG2:1 and IG4:1 are significantly higher than those of hafnium oxide (HfO2).

[0091] 2. The particle sizes of IW2:1, IW3:1, IW4:1, IG2:1, IG3:1, IG4:1 and IOE were detected by DLS particle size analyzer.

[0092] The results are shown in Figure 2. As shown in Figure 2a, the iodized oil emulsion particle size varies, but the emulsions of different particle sizes all show good radiosensitization effect. In the figure, IW21, IW31, IW41, IG21, IG31 and IG41 correspond to IW2:1, IW3:1, IW4:1, IG2:1, IG3:1 and IG4:1, respectively, and the corresponding manner is the same in the other figures. As shown in Figure 2b, the iodized oil emulsion particle size does not change over time (0D is 0 days, 40D is 40 days), and the emulsion has high stability.

[0093] 3. Radiotherapy sensitization experiment 1 (liver cancer)

[0094] C57BL / 6 mice, 6 weeks old; mouse subcutaneous hepatocellular carcinoma (hepa1-6) model; tumor volume 50-100 mm 3 Treatment began with a single intratumoral injection of 50 μL of each drug. Tumor volume was measured every other day after the injection.

[0095] The groups used included blank control, X-ray alone, X-ray combined with hafnium oxide (HfO2), IO, IG2:1, IW4:1 or IG4:1.

[0096] The X-ray irradiation method is: 2 Gy once every other day starting from the 12th day, for a total of three irradiations.

[0097] Correlation test result as shown in Figure 3, the arrow below the abscissa represents that respectively at the 12th day, the 14th day, the 16th day, test mice were carried out to three X-ray irradiations.The result shows, 10+X-ray group and only with no statistically significant difference between the X-ray group.IG4:1, IW4:1 and IG2:1 have obvious sensitization effect to the radiotherapy of liver cancer, are better than the hafnium oxide nanometer radiotherapy sensitizer (about 50nm) used clinically.The iodized oil content is higher, and the effect of radiotherapy sensitization is better.

[0098] 4. Radiotherapy Sensitization Experiment 2 (Breast Cancer)

[0099] BALB / c mice, 6 weeks old; mouse orthotopic mammary carcinoma (4T1) model; tumor volume 50-100 mm 3 Treatment began with a single intratumoral injection of 50 μL of each drug. Tumor volume was measured every other day after the injection.

[0100] The groups used included blank control, X-ray alone, and X-ray combined with 10, IG2:1, IW4:1, or IG4:1.

[0101] The X-ray irradiation method is: 2 Gy once every other day starting from the 12th day, for a total of three irradiations.

[0102] The relevant test results are shown in Figure 4. The arrows below the horizontal axis indicate three X-ray irradiations of the experimental mice on days 12, 14, and 16, respectively. The results showed that there was no statistically significant difference between the IO + X-ray group and the X-ray only group, and that IG4:1 and IW4:1 had a significant sensitization effect on radiotherapy for breast cancer.

[0103] 5. Radiotherapy sensitization experiment 3 (pancreatic cancer)

[0104] BALB / c mice, 6 weeks old; mouse subcutaneous pancreatic cancer model; tumor volume 50-100 mm3 Treatment began with a single intratumoral injection of 25 μL of each drug in each group, followed by a single 6 Gy X-ray irradiation. Tumor volume was measured every other day after the injection.

[0105] Related test results are shown in Figure 5, where the abscissa represents time and the ordinate represents tumor size. Control represents the X-ray group, and Emulsion represents the lipiodol emulsion and X-ray group. The lipiodol emulsion consists of 60% lipiodol, 40% 5% glucose in water, and 5% emulsifier (80% tween 80 + 20% polyoxyethylene hydrogenated castor oil (HCO-40)). As can be seen from the results, lipiodol emulsion has a significant sensitizing effect on pancreatic cancer radiotherapy.

[0106] 6. Radiotherapy sensitization experiment 4 (breast cancer lymph node metastasis)

[0107] BALB / c mice, 6 weeks old; mouse breast cancer footpad model; tumor volume 50-100 mm 3 Treatment began with a single intratumoral injection of 25 μL of each drug in each group. The patients were then irradiated with a single 6 Gy X-ray fraction (both the in situ tumor and the lymph node metastases, i.e., the footpad and thigh root). Following the injection, tumor volume was measured every two or four days (at a later stage).

[0108] The relevant test results are shown in Figure 6. The mice were irradiated with X-rays once on day 8. The results show that iodized oil emulsion IOG has a significant sensitizing effect on radiotherapy of breast cancer. IOG: 20% water for injection, 80% oil phase (40% iodized oil and 60% soybean oil), and 5% emulsifier polyoxyethylene hydrogenated castor oil HCO-40.

[0109] 7. Testing of synergistic effects of radiosensitization and loaded chemical drugs (liver cancer)

[0110] C57 / L mice, 6 weeks old; subcutaneous liver cancer model; volume 50-100 mm 3 Fractionated radiotherapy was initiated; a single dose of 50 μL was injected into the tumor. Tumor volume was measured every other day after the injection.

[0111] The groups used included blank control, X-ray only, and X-ray + IW2:1 loaded cisplatin drug.

[0112] The X-ray irradiation method is: 2 Gy once every other day starting from the 8th day, for a total of three irradiations.

[0113] The relevant test results are shown in Figure 7a, where the arrows below the horizontal axis indicate three X-ray irradiations of the experimental mice on days 8, 10, and 12. As can be seen from the figure, the iodized oil emulsion (IW2:1) loaded with cisplatin has a significant inhibitory effect on mouse tumors.

[0114] 8. Testing of synergistic effects of radiosensitization and loaded chemical drugs (pancreatic cancer)

[0115] C57 / L mice, 6 weeks old; subcutaneous pancreatic cancer model; volume 50-100 mm 3 Radiotherapy was initiated; a single dose of 25 μL was injected into the tumor. Tumor volume was measured every other day after the injection.

[0116] The groups used included blank control, drug-loaded iodized oil emulsion (IOD), X-ray only, pure drug (gemcitabine saline solution, D) + X-ray, iodized oil emulsion (IOE) + X-ray, and drug-loaded iodized oil emulsion (IOD) + X-ray.

[0117] The X-ray irradiation method was: single irradiation of 8 Gy on the 8th day.

[0118] The relevant test results are shown in Figure 7(b), where the arrow below the horizontal axis indicates a single X-ray exposure of the experimental mice on day 8. As can be seen from the figure, lipiodol emulsion can effectively enhance the radiosensitization effect of pancreatic cancer. Furthermore, the drug-loaded lipiodol emulsion further enhances tumor inhibition and achieves radiosensitization and chemotherapy for pancreatic cancer.

[0119] In addition, it can be seen from the whole tumor section (c in Figure 7) that, compared with free gemcitabine, iodized oil emulsion can effectively increase the drug's DNA damage to the tumor, improve the drug's killing effect on the tumor, and achieve chemotherapy sensitization.

[0120] 9. Testing of radiosensitization performance of different tumors

[0121] This example further adopts the following method to test the radiosensitization performance of different tumor cells, as follows:

[0122] Human tumor cell lines (including Hela, PANC-1, A549, HCT116, B16, 143B, or U87) were seeded in 96-well plates (2-4 × 10 3cells). 24 hours later, the culture medium was replaced with lipiodol (30 μg / ml) for 12 hours. The cells were then treated with X-rays (4 Gy for A549 and HCT116 cells, 2 Gy for other cells). Three days later, the absorbance of the supernatant at 450 nm was measured using the CCK assay, revealing that lipiodol enhanced cell radiation damage (cells in the lipiodol-treated group showed a greater killing effect under X-ray irradiation).

[0123] The results, shown in Figure 8, show that lipiodol enhances radiosensitivity in various cancer cell types (from left to right: A549 lung cancer, HeLa cervical cancer, PANC1 pancreatic cancer, HCT116 colon cancer, U87 glioma, B16 melanoma, and 143B osteosarcoma). This significant radiosensitization effect, likely due to the formation of an oil-in-water emulsion when lipiodol is mixed with culture medium, is observed in cellular experiments.

[0124] 10. Test of targeting lymph nodes with lipiodol and imaging

[0125] BALB / C mice, 6 weeks old; footpad breast cancer lymph node metastasis model; 25 μL of drug was injected after tumor lymph node metastasis (approximately 9 days).

[0126] The medications used in each group included: iodized oil (IO) only, IW4:1, and IG4:1.

[0127] Lymph node metastasis of the lipiodol emulsion was observed under CT scan on day 0, immediately after the lipiodol emulsion was injected into the footpad tumor (the arrow points to the popliteal fossa, which is the lymph node location of the lipiodol imaging). After 7 days, the lipiodol emulsion still remained in the in situ tumor and lymph node metastasis. The test results are shown in Figure 9, which shows that both lipiodol and lipiodol emulsion have the ability to target lymph nodes and remain in the lymph nodes for a long time.

[0128] The present invention further verifies the radiosensitization effect of iodized oil emulsion on lymph node metastasis by detecting the volume of lymph nodes, and obtains the result shown in the right figure of Figure 10.

[0129] The groups used included blank control group, X-ray only group, X-ray+IW4:1 group, and X-ray+IG4:1 group.

[0130] The drug injection and X-ray treatment method is as follows: after the tumor lymph node metastasis (about the 9th day), 25 μL of drug is injected and a single 6 Gy X-ray irradiation is performed.

[0131] The present invention further conducted the same experiment using IOG (IOG: 20% water for injection, 80% oil phase (comprising 40% lipiodol and 60% soybean oil), and 5% emulsifier polyoxyethylene hydrogenated castor oil (HCO-40), yielding the results shown in the left figure of Figure 10 . The results demonstrate that lipiodol emulsion inhibits the size of lymph node metastases and improves the radiotherapy clearance of lymph node metastases. Furthermore, the figure demonstrates that iodized oil emulsions, including IOG, IW4:1, and IG4:1, all have a radiotherapy clearance effect on lymph node metastases.

[0132] 11. Intratumoral distribution of iodized oil emulsion and hafnium oxide

[0133] This example further detects the intratumoral distribution of different iodized oil emulsions and hafnium oxide using the following method:

[0134] (1) Mouse orthotopic breast cancer model; tumor volume is approximately 50 mm 3 For drug administration, each group was intratumorally injected with 10 μL of different solutions, including HfO2 or IG4:1.

[0135] (2) Mouse subcutaneous pancreatic cancer model; tumor volume is approximately 50 mm 3 Each group was intratumorally injected with 10 μL of different solutions, including HfO2 or IOE.

[0136] Then, after 4 hours, the tumors were isolated and imaged using a synchrotron-based uCT device (Beijing Synchrotron Radiation Facility).

[0137] Results: Comparison of the intratumoral distribution of hafnium oxide and iodized oil emulsion (IG41, also known as IG4:1) (Figure 11a) and hafnium oxide and iodized oil emulsion (IOE) (Figure 11b) showed that liquid radiotherapy sensitizers have much better diffusion effects than solid nanoparticle sensitizers used clinically, and are more conducive to tumor treatment.

[0138] 12. Iodized oil itself has no therapeutic effect, nor does it have a radiotherapy sensitization effect.

[0139] BALB / c mice, 6 weeks old; mouse subcutaneous breast cancer model; tumor volume 50-100 mm 3 Treatment began with a single intratumoral injection of 50 μL of each drug in each group. Tumor volumes were measured at regular intervals after the injection.

[0140] The groups used included blank control, iodized oil, X-ray group, IO+X-ray group, iohexol+X-ray group

[0141] The X-ray irradiation method was: a single irradiation of 6 Gy on the 8th day.

[0142] The results are shown in Figure 12. When IO (iodized oil) was used alone without X-ray, the tumor size was the same as the control and did not decrease. After radiotherapy, neither iodized oil (IO) nor iohexol had a radiosensitizing effect, which was consistent with the results of the X-ray group.

[0143] 13. Stabilize iodized oil emulsion to improve the safety of iodized oil entering the blood

[0144] Six-week-old C57 mice were injected with 25 μL of each formulation via the tail vein. Groups included blank control, iodized oil (IO), and iodized oil emulsion (IOE). Lung sections were imaged 15 days later.

[0145] The results are shown in Figure 13. IO (iodized oil) entering the blood caused obvious infiltration of the lungs, indicating inflammatory lung damage. The emulsified iodized oil emulsion (IOE) showed good safety in the mouse lungs and did not cause an inflammatory response, which illustrates the advantages of iodized oil emulsion. Industrial Applicability

[0146] The present invention provides a radiotherapy sensitizer, which consists of an oil phase and an aqueous phase, wherein the main component of the oil phase is lipiodol. During research, the present invention discovered that although lipiodol itself does not have a radiotherapy sensitization effect, it can achieve excellent radiotherapy sensitization effects after being prepared into an emulsion. The application of the lipiodol emulsion provided by the present invention has high radiation absorption, high tumor targeting, high tumor diffusion and retention time, as well as good clinical safety. Its effect of reducing tumor volume and extending the life cycle of animals is even better than that of existing hafnium oxide radiotherapy sensitizers, and its safety factor is much higher than that of hafnium oxide, which is of great significance in the field of radiotherapy.

Claims

1. A radiotherapy sensitizer, characterized in that: The radiotherapy sensitizer consists of an oil phase and an aqueous phase; the oil phase includes iodized oil, and the volume percentage of the oil phase in the radiotherapy sensitizer is 30-90%; the volume percentage of the iodized oil in the oil phase is above 30%.

2. A radiotherapy sensitizer, characterized in that: The radiotherapy sensitizer consists of an oil phase, an aqueous phase and a surfactant; the oil phase includes iodized oil, and the volume percentage of the oil phase in the radiotherapy sensitizer is 30-90%.

3. The radiosensitizer according to claim 1 or 2, characterized in that: The oil phase also includes: one or more of vegetable oil, glycerol, octadecyl alcohol or stearic acid; and / or, the aqueous phase includes: one or more of non-ionic contrast agent solution, sodium chloride solution, water, glucose aqueous solution, Ringer's solution, sodium lactate Ringer's solution, acetate Ringer's solution, compound sodium acetate Ringer's solution, sodium bicarbonate Ringer's solution, hydroxyethyl starch, succinylated gelatin, dextran, sugar saline, etc.

4. The radiosensitizer according to claim 2, characterized in that The surfactants include: One or more of nonionic surfactants, anionic surfactants, cationic surfactants, natural surfactants, liposomes and polymers.

5. A radiotherapy drug preparation, characterized in that: The radiotherapy drug preparation comprises: A drug or a prodrug thereof having a tumor therapeutic effect, and the radiosensitizer according to any one of claims 1 to 4.

6. The radiotherapy drug preparation according to claim 5, characterized in that: The drugs having chemotherapeutic effects include any one or more of the following: i) Compounds that affect tissue growth, cell growth or cell differentiation; ii) compounds that induce biological effects; iii) Nanomedicines with the ability to promote radiosensitization or induce cellular changes.

7. The radiotherapy drug preparation according to claim 6, characterized in that: The drugs having tumor therapeutic effects include: One or more of chemotherapeutic drugs, protein drugs, antibody drugs, anti-tumor vascular drugs, anti-tumor targeted drugs or nano drugs.

8. A kit, characterized in that: The kit comprises the radiosensitizer according to any one of claims 1 to 4, or the radiotherapy drug preparation according to any one of claims 5 to 7.

9. The use of iodized oil emulsion as a radiotherapy sensitizer or lymph node tracer; The iodized oil emulsion comprises: Oil phase and water phase; The oil phase comprises iodized oil, and the volume percentage of the oil phase in the iodized oil emulsion is 30-90%; the volume percentage of the iodized oil in the oil phase is above 30%.

10. The use according to claim 9, characterized in that: The oil phase also includes: one or more of vegetable oil, glycerol, octadecyl alcohol or stearic acid; and / or, the aqueous phase includes: one or more of non-ionic contrast agent solution, sodium chloride solution, water, glucose aqueous solution, Ringer's solution, sodium lactate Ringer's solution, acetate Ringer's solution, compound sodium acetate Ringer's solution, sodium bicarbonate Ringer's solution, hydroxyethyl starch, succinylated gelatin, dextran, sugar saline, etc.

11. The use according to claim 9 or 10, characterized in that: The iodized oil emulsion also includes a surfactant.

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