CN-122028933-A - Methods and systems for improved delivery via ultrasound

Abstract

Methods for improving expression of nucleic acid constructs in cells or organs of a subject using sonopore formation and optimization of the ultrasound acoustic energy mechanical index are provided.

Inventors

• Steven. B. Feinstein
• KENNETH GREENBERG
• BARRY CAMPBELL
• Ivan Krivija

Assignees

• 索诺疗法公司

Dates

Publication Date

20260512

Application Date

20240725

Priority Date

20230728

Claims (20)

1. 1. A method of delivering a nucleic acid payload to a target cell of a subject, comprising: a. administering to the subject a nucleic acid construct comprising the nucleic acid payload; b. Administering a sonoactive agent to the subject; c. Applying ultrasonic acoustic energy to the target cells at a first Mechanical Index (MI) of at most 0.4, and D. Applying ultrasonic acoustic energy to the target cells at a second MI greater than 1.3 and up to 2.9.
2. 2. A method of delivering a nucleic acid payload to a target cell of a subject, comprising: a. administering to the subject a nucleic acid construct comprising the nucleic acid payload; b. Administering a sonoactive agent to the subject; c. Applying ultrasonic acoustic energy to the target cells at a first Mechanical Index (MI) of at most 0.4, and D. applying ultrasonic acoustic energy to the target cells at a second MI of at least 2.0.
3. 3. A method of delivering a nucleic acid payload to a target cell of a subject, comprising: a. administering to the subject a nucleic acid construct comprising the nucleic acid payload, wherein the nucleic acid construct is a miniplasmid; b. Administering a sonoactive agent to the subject; c. Applying ultrasonic acoustic energy to the target cells at a first Mechanical Index (MI) of at most 0.4, and D. Applying ultrasonic acoustic energy to the target cells at a second MI greater than 0.4 and up to 2.3.
4. 4. The method of any one of claims 1 or 2, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of greater than 1.5 and up to 2.9.
5. 5. The method of any one of claims 1 or 2, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of greater than 1.8 and at most 2.9.
6. 6. The method of any one of claims 1 or 3, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of at least 2.0.
7. 7. The method of any one of claims 1-3, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of at least 2.2.
8. 8. The method of any one of claims 1 or 2, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of at least 2.4.
9. 9. The method of any one of claims 1 or 23, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of at least 2.6.
10. 10. The method of any one of claims 1 or 2, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of at least 2.9.
11. 11. The method of any one of claims 1 or 2, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of greater than 2.2 and up to 2.9.
12. 12. The method of any one of claims 1 or 2, wherein the ultrasonic acoustic energy is applied to the target cells at the second MI of greater than 2.6 and up to 2.9.
13. 13. The method of any of the preceding claims, wherein the ultrasound transducer that applies the ultrasound acoustic energy to the target cells is in continuous contact with the tissue of the subject and continuously (1) applies the ultrasound acoustic energy to the subject or (2) receives reflected ultrasound acoustic energy from the subject.
14. 14. The method of any one of the preceding claims, wherein the nucleic acid construct is a miniplasmid, wherein the miniplasmid is less than or equal to 500 base pairs in length, excluding an expression cassette.
15. 15. The method of any one of the preceding claims, wherein the nucleic acid construct is administered systemically.
16. 16. The method of any of the preceding claims wherein applying the ultrasonic acoustic energy at the first MI and applying the ultrasonic acoustic energy at the second MI are repeated at least twice.
17. 17. The method of any of the preceding claims wherein the applying the ultrasonic acoustic energy at the first MI and the applying the ultrasonic acoustic energy at the second MI are repeated 4 to 18 times.
18. 18. The method of any of the preceding claims wherein the applying the ultrasonic acoustic energy at the first MI and the applying the ultrasonic acoustic energy at the second MI are repeated 6 to 12 times.
19. 19. The method of any of the preceding claims wherein the applying the ultrasonic acoustic energy at the first MI and the applying the ultrasonic acoustic energy at the second MI are repeated 8 to 10 times.
20. 20. The method of any of the preceding claims wherein the applying the ultrasonic acoustic energy at the first MI and the applying the ultrasonic acoustic energy at the second MI are repeated at least 9 times.

Description

Methods and systems for improved delivery via ultrasound Cross Reference to Related Applications The present application claims the benefit of U.S. provisional patent application No. 63/516,488 filed on day 28 of 7 in 2023 and U.S. provisional patent application No. 63/592,106 filed on day 20 of 10 in 2023 and U.S. provisional patent application No. 63/656,376 filed on day 5 of 6 in 2024, each of which is incorporated herein by reference in its entirety for all purposes. Background Gene therapy has been proposed as a possible method for treating genetic diseases by transfecting functional copies of genes into cells. However, prior art methods of gene therapy using ultrasound or sonopore (sonoporation) suffer from significant drawbacks such as low transfection efficiency and insufficient gene expression, which have hampered the clinical development and commercialization of these methods. There remains a need in the art for effective gene therapy techniques that can transfect genes into cells in an organ or tissue of a subject in a safe, effective, and durable manner. Disclosure of Invention In ultrasound therapy, the Mechanical Index (MI) is a unitless number that measures the power of the ultrasound beam and its potential to cause biological effects in tissue. Sonopore-forming protocols for gene therapy products generally seek to maximize nucleic acid delivery to cells while minimizing the application of ultrasonic energy to cells as much as possible, and in particular seek to maintain the application of ultrasound at a low mechanical index in order to improve the safety and tolerability of cellular procedures. It has been demonstrated that in some cases the application of ultrasound at high mechanical indices induces tissue damage caused by uncontrolled cavitation, inflammatory reactions and vascular damage in the target tissue, while often failing to achieve the goal of ultrasound therapy. The present disclosure provides methods of optimizing delivery of nucleic acid payloads to cells using an ultrasound protocol in combination with elevated mechanical index ultrasound, which remains safe while also significantly increasing delivery of nucleic acid payloads to target cells. As described herein, application of ultrasound at elevated mechanical indices using an alternating mechanical index protocol induces stable vibration and inertial cavitation of the acoustically active agent administered to the subject and results in increased delivery of the nucleic acid payload to the target cells without significantly reducing the safety and tolerability of the procedure within the tissue. Aspects disclosed herein provide a method of delivering a nucleic acid payload to a target cell of a subject, the method comprising administering a nucleic acid construct comprising the nucleic acid payload to the subject, administering a sonoactive agent to the subject, applying ultrasonic acoustic energy to the target cell at a first Mechanical Index (MI) of at most 0.4, and applying ultrasonic acoustic energy to the target cell at a second MI of greater than 1.3 and at most 2.9. Aspects disclosed herein provide a method of delivering a nucleic acid payload to a target cell of a subject, the method comprising administering a nucleic acid construct comprising the nucleic acid payload to the subject, administering a sonoactive agent to the subject, applying ultrasonic acoustic energy to the target cell at a first Mechanical Index (MI) of at most 0.4, and applying ultrasonic acoustic energy to the target cell at a second MI of at least 2.0. Aspects disclosed herein provide a method of delivering a nucleic acid payload to a target cell of a subject, the method comprising administering a nucleic acid construct comprising the nucleic acid payload to the subject, wherein the nucleic acid construct is a microplasma, administering a sonotrode to the subject, applying ultrasonic acoustic energy to the target cell at a first Mechanical Index (MI) of at most 0.4, and applying ultrasonic acoustic energy to the target cell at a second MI of greater than 0.4 and at most 2.3. In some embodiments, ultrasonic acoustic energy is applied to the target cells at a second MI greater than 1.5 and up to 2.9. In some embodiments, ultrasonic acoustic energy is applied to the target cells at a second MI greater than 1.8 and up to 2.9. In some embodiments, ultrasonic acoustic energy is applied to the target cells at a second MI of at least 2.0. In some embodiments, ultrasonic acoustic energy is applied to the target cells at a second MI of at least 2.2. In some embodiments, ultrasonic acoustic energy is applied to the target cells at a second MI of at least 2.4. In some embodiments, ultrasonic acoustic energy is applied to the target cells at a second MI of at least 2.6. In some embodiments 3, ultrasonic acoustic energy is applied to the target cells at a second MI of at least 2.9. In some embodiments, ultrasonic acoustic energy is applied to the target