# US 12,109,440 B2 · Compositions, methods and systems for gas vesicle based cavitation

> Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt

- Number: US 12,109,440 B2
- Document ID: US12109440B2
- Status: Granted
- Kind: B2
- Application number: 17/816,373
- Filed: 2022-07-29
- Published: 2023-05-04
- Granted: 2024-10-08
- Earliest claimed: 2019-03-28
- Assignees: California Institute of Technology
- Applicants: California Institute of Technology
- Inventors: Avinoam D. Bar-Zion; Atousa Nourmahnad; David Maresca; Mikhail Shapiro
- Web page: https://patentgrep.com/patent/US12109440B2

## CPC

- A61N 7/00: Ultrasound therapy (lithotripsy A61B17/22, A61B17/225; massage using supersonic vibration A61H23/00 {; using ultrasound for introducing media into the body A61M37/0092})
- A61M 37/0092: {using ultrasonic, sonic or infrasonic vibrations, e.g. phonophoresis}
- A61B 2090/374: {NMR or MRI}
- A61N 2007/0004: {Applications of ultrasound therapy}
- A61N 2007/0039: {using microbubbles}
- A61N 2007/0052: {using the same transducer for therapy and imaging}
- A61N 2007/0082: {Scanning transducers}
- A61N: ELECTROTHERAPY; MAGNETOTHERAPY; RADIATION THERAPY; ULTRASOUND THERAPY (measurement of bioelectric currents A61B; surgical instruments, devices or methods for transferring non-mechanical forms of energy to or from the body A61B18/00; anaesthetic apparatus in general A61M; incandescent lamps H01K; infrared radiators for heating H05B)
- A61M: DEVICES FOR INTRODUCING MEDIA INTO, OR ONTO, THE BODY (introducing media into or onto the bodies of animals A61D7/00; means for inserting tampons A61F13/26; devices for administering food or medicines orally A61J; containers for collecting, storing or administering blood or medical fluids A61J1/05); DEVICES FOR TRANSDUCING BODY MEDIA OR FOR TAKING MEDIA FROM THE BODY (surgery A61B; chemical aspects of surgical articles A61L); DEVICES FOR PRODUCING OR ENDING SLEEP OR STUPOR {(Electrotherapy, e.g. producing anaesthesia by the use of alternating or intermittent currents A61N1/36021)}
- A61B: DIAGNOSIS; SURGERY; IDENTIFICATION (usefulness limited to only animals A61D)

## Abstract

The system and process of therapeutic and effective cavitation by using ultrasound to collapse gas vesicles as well as cavitate the bubbles produced from the collapsed gas vesicles. Therapeutic effect includes, but is not limited to lysing cells by cavitation. The cells expressing the gas vesicles can optionally be used as delivery cells to preform tasks such as transporting the gas vesicles into deep tissue areas, releasing compounds at the cavitation site, and more. The gas vesicles can optionally be modified to facilitate getting the bubbles near the cavitation targets by functionalizing the gas vesicles.

## Claims

### Claim 1

An ultrasound pressure-based method comprising:
delivering a gas vesicle (GV) to a target site, the GV having a GV type collapse threshold pressure; setting an ultrasound generation system to produce ultrasound pulses having a positive pressure set above the GV type collapse threshold pressure, a negative pressure and at least one of: a duty cycle above 0.2% or a mechanical index above 2.8; and applying the ultrasound pulses to the target site, thus: collapsing the GV by the positive pressure of the ultrasound pulses, thereby forming gas bubbles; and
driving a cavitation of the gas bubbles by the negative pressure wherein the gas vesicle comprises a variant GvpC engineered to comprise binding tags presented on the GV and configured to bind to a target molecule on a cavitation target.

### Claim 2 (depends on 1)

The method of claim 1, further comprising imaging the target site before applying the ultrasound pulses.

### Claim 3 (depends on 2)

The method of claim 2, wherein the imaging comprises one of ultrasound imaging or MRI.

### Claim 4 (depends on 3)

The method of claim 3, wherein the imaging comprising using the GV as a contrast agent.

### Claim 5 (depends on 2)

The method of claim 2, further comprising imaging the target site after applying the ultrasound pulses.

### Claim 6 (depends on 1)

The method of claim 1, wherein the ultrasound pulses are applied with a center frequency no greater than 2.5 MHz.

### Claim 7 (depends on 6)

The method of claim 6, wherein the ultrasound pulses have a mechanical index above 0.12.

### Claim 8 (depends on 1)

The method of claim 1, wherein the GV is a plurality of GVs.

### Claim 9

A method of applying a mechanical stress to a cavitation target in a target site, the method comprising: delivering to the target site containing the cavitation target one or more gas vesicles (GVs) having a GV type collapse threshold, the delivering positioning the GVs in proximity of the cavitation target, and applying ultrasound pulses to the target site after the delivering, the ultrasound pulses selected to both collapse the GV type by having a positive pressure above the GV type collapse threshold and to drive the cavitation of bubbles created by the GVs after collapse by negative pressure of the ultrasound pulses, the cavitated bubbles applying the mechanical stress to the cavitation target; wherein the gas vesicle comprises a variant GvpC engineered to comprise binding tags presented on the GVs and configured to bind to a target molecule on the cavitation target.

### Claim 10 (depends on 9)

The method of claim 9, wherein the cavitation target is an inorganic target.

### Claim 11 (depends on 9)

The method of claim 9, wherein the mechanical stress is selected from the group consisting of destroying, damaging, pushing, pinching, propelling, mixing, and deforming.

### Claim 12 (depends on 9)

The method of claim 9, further comprising imaging the target site at a first time to verify delivery of the GVs at the target site and imaging the target site at a second time to verify an effect of the cavitating of the bubbles.

### Claim 13 (depends on 1)

The method of claim 1, being a method of lysing a target cell of a target tissue at the target site, further comprising lysing the target cell by the cavitated bubbles.

### Claim 14 (depends on 1)

The method of claim 1, wherein the ultrasound pulse has a center frequency selected to be less than 2.5 MHz.

### Claim 15 (depends on 14)

The method of claim 14, wherein the cavitation target is a target tissue and the GV are functionalized to attach to the target tissue.

### Claim 16 (depends on 14)

The method of claim 14, wherein the cavitation target is a target tissue and the delivering the GV to the target tissue comprises expressing the GV in a delivery cell that is delivered to the target tissue.

### Claim 17 (depends on 14)

The method of claim 14, wherein a target cell is within a target tissue and the GV contains a payload compound that has a therapeutic effect on the target tissue.

### Claim 18 (depends on 1)

The method of claim 1, wherein the delivering the GV to the target site comprises delivering a delivery cell containing the GV, and wherein the cavitated bubbles lyse a target cell by lysing the delivery cell.

### Claim 19 (depends on 18)

The method of claim 18, wherein the delivery cell expresses the GV.

### Claim 20 (depends on 18)

The method of claim 18, wherein the delivery cell contains a hybrid cluster.

### Claim 21 (depends on 1)

The method of claim 1, being a method of delivering a payload compound to a target tissue at the target site, wherein the delivering the GV to the target site comprises delivering a delivering cell configured to express the payload compound, the delivering performed for a time and under conditions allowing expression of the GVs in the delivery cell, and wherein the cavitated bubbles lyse the delivery cell and deliver the payload compound to the tissue.

### Claim 22 (depends on 21)

The method of claim 21, wherein the payload compound is a protein expressed by the delivery cell.

### Claim 23 (depends on 21)

The method of claim 21, wherein the allowing the expression of the GV in the delivery cell occurs before the delivering the delivery cell.

### Claim 24 (depends on 21)

The method of claim 21, further comprising ultrasound imaging or MRI imaging the delivery cell at the target site.

### Claim 25 (depends on 21)

The method of claim 21, wherein the payload compound is a therapeutic substance.

### Claim 26 (depends on 1)

The method of claim 1, wherein the cavitation target is a target cell within the target site and the engineered GVPc variant comprises a cell targeting moiety specific for the target cell.

### Claim 27 (depends on 26)

The method of claim 26, wherein the cell targeting moiety is a receptor-targeting peptide RGD.

### Claim 28 (depends on 26)

The method of claim 26 wherein the cell targeting moiety is specific for macrophages.

### Claim 29 (depends on 28)

The method of claim 28, wherein the cell targeting moiety is a CD47 or R8 moiety.

### Claim 30 (depends on 8)

The method of claim 8, wherein the plurality of GVs comprises two or more populations of GVs each having different collapse pressures.

### Claim 31 (depends on 9)

The method of claim 9, wherein the cavitation target is a target cell within the target site and the engineered GVPc variant comprises a cell targeting moiety specific for the target cell.

### Claim 32 (depends on 31)

The method of claim 31, wherein the cell targeting moiety is a receptor-targeting peptide RGD.

### Claim 33 (depends on 32)

The method of claim 32 wherein the cell targeting moiety is specific for macrophages.

### Claim 34 (depends on 33)

The method of claim 33, wherein the cell targeting moiety is a CD47 or R8 moiety.

### Claim 35 (depends on 9)

The method of claim 9, wherein the one or more gas vesicles (GVs) comprise a plurality of GVs each having a GV type collapse threshold comprise two or more populations of GVs each having different collapse pressures.