Application of oncolytic virus combined with high-intensity focused ultrasound to treatment of pelvic malignant tumor

Abstract

The present invention provides the use of a treatment system comprising an oncolytic virus drug delivery device configured with an oncolytic virus and a high intensity focused ultrasound (HIFU) device in the treatment of pelvic malignancies. Clinical experiments prove that combined application of the oncolytic virus and high-intensity focused ultrasound has an excellent treatment effect on the pelvic malignant tumor, growth and invasion of the pelvic malignant tumor of a subject are effectively inhibited, and almost no adverse reaction occurs. The invention provides a more positive and effective scheme for pelvic malignant tumor treatment.

Description

Technical Field

[0001] The present invention relates to the field of medical technology, and in particular to the application of oncolytic virus combined with high-intensity focused ultrasound (HIFU) in the treatment of pelvic malignant tumors. Background Art

[0002] High-Intensity Focused Ultrasound (HIFU) is a new non-invasive tumor treatment technology that uses the inherent characteristics of ultrasound to focus low-energy ultrasound into solid tumors, forming instantaneous high temperatures, while exerting the cavitation effect and mechanical effect of ultrasound, causing tumor cells to degenerate and necrotize, achieving the purpose of directly killing tumors; at the same time, the inactivated tumor tissue acts as a tumor antigen, stimulating the body to produce anti-tumor antibodies, which has a subsequent immune anti-tumor effect. Oncolytic viruses are an important class of gene therapy drugs with advantages such as more precise targets, better sustained effects, and fewer side effects.

[0003] Pelvic malignant tumors are currently a high-morbidity disease in clinical practice, which seriously threatens the life safety of patients. With the continuous changes in people's lifestyles in recent years, the incidence of pelvic malignant tumors has become increasingly higher. In particular, pelvic metastasis occurs after surgical resection and adjuvant chemotherapy of colon cancer and ovarian cancer, which seriously affects the quality of life and health of patients. Therefore, it is necessary to find an active and effective comprehensive treatment plan. Summary of the invention

[0004] The object of the present invention is to provide a treatment system comprising an oncolytic virus delivery device configured with oncolytic virus and a high-intensity focused ultrasound device for use in treating pelvic malignant tumors.

[0005] In a first aspect of the present invention, a treatment system is provided for use in preparing a product for treating pelvic malignant tumors, wherein the system comprises an oncolytic virus delivery device configured with an oncolytic virus and a high-intensity focused ultrasound device.

[0006] In another preferred example, the oncolytic virus administration device is used to administer the configured oncolytic virus to a subject.

[0007] In another preferred embodiment, the oncolytic virus is administered by intratumoral injection.

[0008] In another preferred embodiment, the oncolytic virus is formulated in a unit dosage form containing the oncolytic virus.

[0009] In another preferred embodiment, the unit dosage form containing the oncolytic virus is a lyophilized agent or a liquid agent, preferably a liquid agent.

[0010] In another preferred embodiment, the oncolytic virus drug delivery device is configured with one or more (such as at least two) unit dosage forms containing oncolytic viruses, preferably 4-10.

[0011] In another preferred embodiment, the oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

[0012] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0013] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0014] In another preferred embodiment, the oncolytic virus is administered at a rate of 0.5×10 12 VP to 2.5×10 12 Dosage administration of VP.

[0015] In another preferred embodiment, the high-intensity focused ultrasound device is used to generate high-intensity focused ultrasound for ablating tumor lesions in a subject.

[0016] In another preferred embodiment, the high-intensity focused ultrasound device is a JC200 focused ultrasound tumor treatment system (Haifu, Chongqing, China).

[0017] In another preferred embodiment, the single treatment time of the high-intensity focused ultrasound device is 200-2000 seconds, and the average treatment power is 72-400 watts.

[0018] In another preferred embodiment, the pelvic malignant tumor is a solid pelvic malignant tumor with a good vocal channel.

[0019] In another preferred embodiment, the pelvic malignant tumors include pelvic in situ tumors and pelvic metastatic tumors.

[0020] In another preferred embodiment, the pelvic malignancy is selected from the following group: pelvic metastasis of colon cancer, pelvic metastasis of ovarian cancer, or a combination thereof.

[0021] In another preferred embodiment, the "treatment of pelvic malignant tumors" refers to:

[0022] (i) inhibiting in situ tumor growth;

[0023] (ii) inhibiting in situ tumor invasion and / or metastasis;

[0024] (iii) inhibiting the growth of metastatic tumors;

[0025] (iv) inhibiting the invasion of metastatic tumors; and / or

[0026] (v) Prolong the survival of the subject.

[0027] In another preferred embodiment, the subject is a human or a non-human mammal.

[0028] A second aspect of the present invention provides a system for treating pelvic malignant tumors, the system comprising:

[0029] (a) an oncolytic virus administration module, which is used to administer an oncolytic virus to a subject, wherein the oncolytic virus administration module is configured with one or more (e.g., at least two) unit dosage forms containing an oncolytic virus, preferably 4-10;

[0030] (b) a high-intensity focused ultrasound module, the high-intensity focused ultrasound module being used to generate high-intensity focused ultrasound to ablate tumor lesions in a subject; and

[0031] (c) a control module, wherein the control module is used to control the system to operate in the order of (a) oncolytic virus administration module (b) → high-intensity focused ultrasound module, or (b) high-intensity focused ultrasound module → (a) oncolytic virus administration module, or to control (a) and (b) to operate simultaneously.

[0032] In another preferred example, the control module is used to control the system to operate in the order of (b) high-intensity focused ultrasound module → (a) oncolytic virus administration module.

[0033] In another preferred embodiment, the oncolytic virus administration module is set to be 0.5×10 12 VP to 2.5×10 12 Dosage administration of VP.

[0034] In another preferred embodiment, the unit dosage form of the oncolytic virus contains 5×10 11 vp / 0.5ml dose of oncolytic virus.

[0035] In another preferred embodiment, the oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

[0036] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0037] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0038] In another preferred embodiment, the high-intensity focused ultrasound module comprises a high-intensity focused ultrasound device, and the device is selected from: JC200 focused ultrasound tumor treatment system (Chongqing Haifu, China).

[0039] In another preferred embodiment, the high-intensity focused ultrasound module is set to a single treatment time of 200-2000 seconds and an average treatment power of 72-400 watts.

[0040] In another preferred embodiment, the system further comprises: (d) an efficacy evaluation module, which is used to evaluate the therapeutic effect of the subject's pelvic malignant tumor.

[0041] In another preferred embodiment, the pelvic malignant tumor is a solid pelvic malignant tumor with a good vocal channel.

[0042] In another preferred embodiment, the pelvic malignant tumors include pelvic in situ tumors and pelvic metastatic tumors.

[0043] In another preferred embodiment, the pelvic malignancy is selected from the following group: pelvic metastasis of colon cancer, pelvic metastasis of ovarian cancer, or a combination thereof.

[0044] The third aspect of the present invention provides a method for treating pelvic malignant tumors, the method comprising using the system of the second aspect of the present invention to treat a subject in need thereof.

[0045] In another preferred embodiment, the subject in need is a human or a non-human mammal.

[0046] In another preferred embodiment, the non-human mammals include monkeys, gorillas, cows, pigs, dogs, sheep, rabbits or mice.

[0047] In another preferred embodiment, the subject in need suffers from pelvic malignant tumors, which include pelvic in situ tumors and pelvic metastatic tumors.

[0048] In another preferred embodiment, the pelvic malignancy is selected from the following group: pelvic metastasis of colon cancer, pelvic metastasis of ovarian cancer, or a combination thereof.

[0049] In another preferred embodiment, the method further comprises administering supportive therapy drugs and / or chemotherapy drugs for treating tumors to a subject in need thereof.

[0050] In another preferred embodiment, the supportive treatment drugs include: antibiotics, analgesics, nutritional drugs or a combination thereof.

[0051] In another preferred embodiment, the chemotherapy drug is selected from irinotecan, capecitabine, oxaliplatin, S-1, bevacizumab or a combination thereof.

[0052] It should be understood that within the scope of the present invention, the above-mentioned technical features of the present invention and the technical features specifically described below (such as embodiments) can be combined with each other to form a new or preferred technical solution. Due to space limitations, they will not be described one by one here. BRIEF DESCRIPTION OF THE DRAWINGS

[0053] Figure 1 Shown are the CT examination results of the pelvic lesion in Case 1 before and after high-intensity focused ultrasound (HIFU) combined with oncolytic virus therapy. DETAILED DESCRIPTION

[0054] After extensive and in-depth research, the inventor unexpectedly discovered for the first time that oncolytic virus (especially recombinant human adenovirus type 5 (Ancore)) combined with high-intensity focused ultrasound (HIFU) has an excellent therapeutic effect on pelvic malignant tumors. Clinical experiments have shown that the combined use of recombinant human adenovirus type 5 and high-intensity focused ultrasound (HIFU) can effectively inhibit the growth of pelvic malignant tumors. On this basis, the present invention was completed.

[0055] Recombinant human adenovirus type 5 injection

[0056] The recombinant human adenovirus type 5 injection used in the present invention, with the trade name of Ankorui, is the first oncolytic virus therapeutic drug approved by the China Food and Drug Administration and launched on the market in 2006. It is an oncolytic adenovirus obtained by genetically recombining human adenovirus type 5 (Ad5) using genetic engineering technology. It mainly deletes the E1B-55kDa (i.e., 55kD in the E1B region of type 5 adenovirus, encoded by the E1B-55k gene) and E3 region gene fragments of human adenovirus type 5, and has the characteristic of specific replication in tumor cells, which ultimately leads to oncolysis.

[0057] The oncolytic mechanism of oncolytic viruses is constantly updated with the progress of tumor immunity research. It was previously believed that because some viruses are naturally tropistic for tumors, they can selectively infect tumor cells and lyse tumor cells to kill the tumor. As the research continues to deepen, it has been found that oncolytic viruses not only have the ability to directly lyse tumors, but also can improve the tumor microenvironment in an immunosuppressive state, turning "cold" tumors into "hot" tumors, thereby activating innate immunity and adaptive immunity, so that the body's immune system that was originally evaded by the tumor is mobilized to attack the tumor.

[0058] Adenovirus is a type of pathogen that has been studied in depth. Since the 1960s, adenovirus has been used as a gene expression vector and has been widely used in the field of molecular biology. It is the most commonly used viral vector for gene therapy. Adenovirus vectors have many characteristics that other vectors do not have. Various types of adenovirus vectors have been successfully constructed and applied in engineering.

[0059] Adenovirus is a DNA virus without an envelope. It is composed of 252 capsomers arranged in an icosahedral pattern and is a particle of 70 to 90 nm. The capsid contains a linear double-stranded DNA molecule with an inverted repeat sequence of about 100 bp at each end. Since the 5' end of each DNA chain has a relative molecular mass of 55×10 3 The protein molecules of Da are covalently bound to form a circular structure of double-stranded DNA.

[0060] The adenovirus E3 region gene is an early transcription unit of the adenovirus genome. Its promoter is activated by the E1A protein and then transcribed into protein. The E3 protein mainly destroys the host's immune defense mechanism, protecting the adenovirus from attack by the immune system in the host.

[0061] The recombinant human adenovirus type 5 used in the present invention has been modified, and after the E3 region gene is deleted, it only has the ability to replicate in the host cell without damaging the host's immune system.

[0062] High-intensity focused ultrasound (HIFU)

[0063] High Intensity Focused Ultrasound (HIFU) is an emerging technology that uses ultrasound as an energy source under the guidance and monitoring of images, and utilizes its penetrability and focusability to focus low-energy ultrasound waves outside the body on the target area inside the human body to form a focal area. The dense ultrasound energy in the focal area can cause the temperature of the local tissue (target area) to rise rapidly in a very short time, and then coagulative necrosis (ablation) occurs, thereby achieving the purpose of treatment without damaging normal structures outside the target area.

[0064] The energy of focused ultrasound in the non-treatment area it passes through is not enough to cause tissue damage. At its focal point, due to the high sound intensity, the thermal effect of ultrasound causes the temperature of the tissue there to rise instantly to 65-100°C, resulting in protein denaturation and coagulative necrosis of tissue cells; at the same time, through the cavitation effect of ultrasound, the interstitial fluid, intercellular fluid and intracellular gas molecules form bubbles under the positive and negative pressure phases of ultrasound, which then shrink and expand until they finally burst, and the energy generated causes cell damage and necrosis. The shape and size of the focused ultrasound focal area, as well as the effect and reaction of the tissue on ultrasound, play a decisive role in the depth of ultrasound treatment, the range of tissue damage and the degree of damage. Therefore, by setting the parameters of the ultrasonic transducer, the purpose of targeted destruction of lesions can be achieved without damaging the tissues around the treatment target, thereby achieving the goal of non-invasive treatment.

[0065] In a preferred embodiment of the present invention, the high-intensity focused ultrasound used is the JC200 focused ultrasound tumor treatment system (Haifu, Chongqing, China).

[0066] System of the present invention

[0067] In one aspect of the present invention, a system for treating pelvic malignancies is provided, the system comprising:

[0068] (a) an oncolytic virus administration module, which is used to administer an oncolytic virus to a subject, wherein the oncolytic virus administration module is configured with one or more (e.g., at least two) unit dosage forms containing an oncolytic virus, preferably 4-10;

[0069] (b) a high-intensity focused ultrasound module, the high-intensity focused ultrasound module being used to generate high-intensity focused ultrasound to ablate tumor lesions in a subject; and

[0070] (c) a control module, wherein the control module is used to control the system to operate in the order of (a) oncolytic virus administration module → (b) high-intensity focused ultrasound module, or (b) high-intensity focused ultrasound module → (a) oncolytic virus administration module, or to control (a) and (b) to operate simultaneously.

[0071] In another preferred embodiment, the oncolytic virus administration module is set to be 0.5×10 12 VP to 2.5×10 12 Dosage administration of VP.

[0072] In another preferred embodiment, the unit dosage form of the oncolytic virus contains 5×10 11 vp / 0.5ml dose of oncolytic virus.

[0073] In another preferred embodiment, the oncolytic virus refers to a recombinant human adenovirus type 5 or a derivative having the same function, preferably, a recombinant human adenovirus type 5.

[0074] In another preferred embodiment, the oncolytic virus is a recombinant human adenovirus type 5 injection with the trade name Ancore.

[0075] In another preferred embodiment, in the genome of the oncolytic virus, the E1B-55kDa and E3 region gene fragments of human adenovirus type 5 are deleted.

[0076] In another preferred embodiment, the high-intensity focused ultrasound module comprises a high-intensity focused ultrasound device, and the device is selected from: JC200 focused ultrasound tumor treatment system (Chongqing Haifu, China).

[0077] In another preferred embodiment, the high-intensity focused ultrasound module is set to a single treatment time of 200-2000 seconds and an average treatment power of 72-400 watts.

[0078] It should be understood that the selection of the high-intensity focused ultrasound device in the high-intensity focused ultrasound module, and the treatment parameters (including treatment time, power, etc.) may vary according to the type, volume, severity, lesion location, etc. of the tumor. The therapeutically effective amount of the oncolytic virus may also vary with the mode of administration and the severity of the tumor. The preferred device selection and parameter setting and the effective amount can be determined by a person of ordinary skill in the art based on various factors (e.g., through clinical trials). The factors include, but are not limited to: pharmacokinetic parameters such as bioavailability, metabolism, half-life, etc.; the severity of the tumor, the patient's weight, the patient's immune status, the route of administration, etc. These are all within the skill range of skilled physicians.

[0079] As used herein, the term "effective amount" or "effective dose" refers to an amount that can produce a function or activity on humans and / or animals and / or cells and can be accepted by humans and / or animals.

[0080] As used herein, the term "unit dosage form" refers to the preparation of an oncolytic virus into a dosage form required for a single administration for the convenience of administration, including but not limited to various solid doses (such as tablets), liquids, capsules, and sustained-release agents. In the present invention, the unit dosage form of the recombinant human adenovirus type 5 is generally a liquid.

[0081] In a preferred embodiment of the present invention, the system comprises:

[0082] (a) Recombinant human adenovirus type 5 injection (Ankorui) drug delivery module, wherein the module is configured with 4 Ankorui unit dosage forms, with a specification of 5×10 11 vp / 0.5ml;

[0083] (b) a high-intensity focused ultrasound module, wherein the module is set to have an average treatment power of 141 W, a treatment irradiation time of 400 seconds, and a total treatment energy of 56390 J; and

[0084] (c) a control module, wherein the control module is used to control the system to operate in the order of (b) high-intensity focused ultrasound module→(a) recombinant human adenovirus type 5 injection (Ankorui) drug delivery module.

[0085] Therapeutic applications

[0086] In another aspect of the present invention, a method for treating pelvic malignancies is provided, the method comprising using the system of the second aspect of the present invention to treat a subject in need thereof.

[0087] In another preferred embodiment, the subject in need suffers from pelvic malignancies, including pelvic in situ tumors and pelvic metastatic tumors. In another preferred embodiment, the pelvic malignancies are selected from the group consisting of pelvic metastasis of colon cancer, pelvic metastasis of ovarian cancer, or a combination thereof.

[0088] In addition, the method may further include administering supportive therapy drugs and / or chemotherapeutic drugs for treating tumors to a subject in need. The supportive therapy drugs include (but are not limited to): antibiotics, analgesics, nutritional drugs or a combination thereof. The chemotherapeutic drugs include (but are not limited to): irinotecan, capecitabine, oxaliplatin, tegafur, bevacizumab or a combination thereof.

[0089] The method of the present invention is very effective for primary or metastatic solid pelvic malignancies with good vocal tracts.

[0090] The main advantages of the present invention are:

[0091] (1) Combining two anti-tumor treatment methods with different mechanisms of action, namely HIFU and local injection of oncolytic viruses, has enhanced the therapeutic effect of pelvic malignancies.

[0092] (2) Both HIFU and local injection of oncolytic viruses have mild adverse reactions and do not affect the normal progress of systemic anti-tumor treatments such as chemotherapy and immunotherapy.

[0093] The present invention will be further described below in conjunction with specific examples. It should be understood that these examples are only used to illustrate the present invention and are not intended to limit the scope of the present invention. Unless otherwise specified, the reagents, methods and equipment used in the present invention are conventional reagents, methods and equipment in the art. Unless otherwise specified, the materials and reagents used in the embodiments of the present invention are all commercially available products. The experimental methods in the following examples that do not specify specific conditions are usually based on conventional conditions, such as Sambrook et al., Molecular Cloning: Laboratory Manual (New York: Cold Spring Harbor Laboratory Press, 1989) The conditions described, or according to the conditions recommended by the manufacturer.

[0094] Unless otherwise indicated, percentages and parts are by weight.

[0095] The efficacy evaluation criteria for pelvic malignant tumors in the present invention are:

[0096]

[0097] Example 1

[0098] Clinical treatment experiment of case 1

[0099] (1) Case introduction

[0100] Patient Chen XX, female, 54 years old. In June 2015, she underwent "radical right hemicolon cancer resection" due to "ascending colon cancer". After the operation, it was shown that "ulcerative adenocarcinoma, moderately differentiated, infiltrating the entire layer of the intestinal wall to the extraserosa fibrofatty tissue, and cancer tissue invading the nerves was visible". After the operation, adjuvant chemotherapy (oxaliplatin combined with Segalovirus) was performed for 6 months. In January 2017, a follow-up examination revealed left lung metastasis, and a left lower lobectomy was performed. Postoperative pathology showed metastatic intestinal adenocarcinoma, and irinotecan combined with capecitabine chemotherapy was performed for 6 months after the operation. In January 2018, a huge pelvic mass was found after the follow-up examination. The biopsy pathology showed adenocarcinoma, and combined with immunohistochemistry, it suggested a colonic origin. From January 2018 to September 2019, chemotherapy combined with targeted therapy and oral anti-vascular therapy were continuously received, and the best therapeutic effect was stable disease (SD).

[0101] (2) Treatment options

[0102] In September 2019, the patient received one cycle of high-intensity focused ultrasound (HIFU) treatment for pelvic masses combined with local injection of oncolytic virus (one cycle refers to one HIFU treatment and one subsequent local injection of recombinant human adenovirus type 5 injection (Ankerui)). The average power of high-intensity focused ultrasound (HIFU) treatment was 141W, the treatment irradiation time was 400 seconds, and the total treatment energy was 56390J; 2ml (5*10^11VP / 0.5ml / tube*4 tubes) of recombinant human adenovirus type 5 injection (Ankerui) was injected locally into the lesion.

[0103] (3) Treatment outcomes

[0104] The results showed that compared with the (HIFU treatment combined with local injection of oncolytic virus) Figure 1 Right), after combined treatment, the patient's abdominal pain and bloating symptoms were alleviated, and the maximum diameter of the tumor was significantly reduced ( Figure 1 Left), it was reduced from 17.6cm before treatment to 11.5cm; the efficacy evaluation was PR (partial remission).

[0105] In addition, the hematological examinations of the patients during the treatment showed that there were no abnormal fluctuations in the total white blood cell count and the cell counts and classification ratios of granulocytes, lymphocytes and monocytes in the blood images before and after treatment, and no obvious adverse reactions were found (Table 1).

[0106] Table 1 Results of hematological examinations before and after treatment

[0107] The above results show that the use of high-intensity focused ultrasound (HIFU) therapy combined with local injection of oncolytic virus therapy not only has significant therapeutic effect but also has good safety.

[0108] All documents mentioned in the present invention are cited as references in this application, just as each document is cited as reference individually. In addition, it should be understood that after reading the above teachings of the present invention, those skilled in the art can make various changes or modifications to the present invention, and these equivalent forms also fall within the scope defined by the claims attached to this application.

Claims

1. Use of a therapeutic system in the preparation of a product for treating pelvic malignant tumors, the system comprising an oncolytic virus delivery device configured with an oncolytic virus and a high-intensity focused ultrasound device.

2. The use according to claim 1, characterized in that The oncolytic virus administration device is used to administer the configured oncolytic virus to a subject.

3. The use according to claim 1, characterized in that The oncolytic virus is configured in a unit dosage form containing the oncolytic virus.

4. The use according to claim 1, characterized in that The oncolytic virus was injected at a rate of 0.5×10 12 VP to 2.5×10 12 Dosage administration of VP.

5. The use according to claim 1, characterized in that The high-intensity focused ultrasound device is used to generate high-intensity focused ultrasound for ablating tumor lesions in a subject.

6. The use according to claim 1, characterized in that The single treatment time of the high-intensity focused ultrasound device is 200-2000 seconds, and the average treatment power is 72-400 watts.

7. The use according to claim 1, characterized in that The pelvic malignancy is selected from the group consisting of pelvic metastasis of colon cancer, pelvic metastasis of ovarian cancer, or a combination thereof.

8. The use according to claim 1, characterized in that The term "treatment of pelvic malignant tumors" refers to: (i) inhibiting in situ tumor growth; (ii) inhibiting in situ tumor invasion and / or metastasis; (iii) inhibiting the growth of metastatic tumors; (iv) inhibiting the invasion of metastatic tumors; and / or (v) Prolong the survival of the subject.

9. A system for treating pelvic malignant tumors, characterized in that: The system comprises: (a) an oncolytic virus administration module, which is used to administer an oncolytic virus to a subject, wherein the oncolytic virus administration module is configured with one or more (e.g., at least two) unit dosage forms containing an oncolytic virus, preferably 4-10; (b) a high-intensity focused ultrasound module, the high-intensity focused ultrasound module being used to generate high-intensity focused ultrasound to ablate tumor lesions in a subject; and (c) a control module, wherein the control module is used to control the system to operate in the order of (a) oncolytic virus administration module (b) → high-intensity focused ultrasound module, or (b) high-intensity focused ultrasound module → (a) oncolytic virus administration module, or to control (a) and (b) to operate simultaneously.

10. The system according to claim 9, characterized in that The system further comprises: (d) an efficacy evaluation module, which is used to evaluate the therapeutic effect of the subject's pelvic malignant tumor.

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