# US 10,864,161 B2 · Systems and methods for targeted breast cancer therapies

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- Number: US 10,864,161 B2
- Document ID: US10864161B2
- Status: Granted
- Kind: B2
- Application number: 16/159,550
- Filed: 2018-10-12
- Published: 2019-04-18
- Granted: 2020-12-15
- Earliest claimed: 2017-10-13
- Assignees: American University of Sharjah
- Applicants: American University of Sharjah
- Inventors: Ghaleb Husseini; Mohammad Al-Sayah; Amal Elsadig
- Web page: https://patentgrep.com/patent/US10864161B2

## CPC

- A61K 9/1271: Non-conventional liposomes, e.g. PEGylated liposomes or liposomes coated or grafted with polymers (liposomes as conjugates {A61K47/6911})
- A61K 31/704: attached to a condensed carbocyclic ring system, e.g. sennosides, thiocolchicosides, escin, daunorubicin {(digitoxin A61K31/7048)}
- A61K 41/0028: {Disruption, e.g. by heat or ultrasounds, sonophysical or sonochemical activation, e.g. thermosensitive or heat-sensitive liposomes, disruption of calculi with a medicinal preparation and ultrasounds}
- A61K 47/6849: {the antibody targeting a receptor, a cell surface antigen or a cell surface determinant}
- A61K 47/6855: {the tumour determinant being from breast cancer cell}
- A61K 47/6913: {the liposome being modified on its surface by an antibody}
- A61M 37/0092: {using ultrasonic, sonic or infrasonic vibrations, e.g. phonophoresis}
- A61P 35/00: Antineoplastic agents
- A61K: PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES (devices or methods specially adapted for bringing pharmaceutical products into particular physical or administering forms A61J3/00; chemical aspects of, or use of materials for deodorisation of air, for disinfection or sterilisation, or for bandages, dressings, absorbent pads or surgical articles A61L; soap compositions C11D)
- 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)}
- A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS

## Abstract

Systems and methods for producing liposomes, including control liposomes and immunoliposomes targeting breast cancer are provided. Systems and methods for treating breast cancer, using targeted immunoliposomes produced according to various methods are also disclosed herein. For example, trastuzumab-conjugated immunoliposomes may be used to deliver chemotherapeutic agents to breast cancer tissues for the treatment of breast cancer. Systems and methods for actuating liposomes using ultrasound are also disclosed, such as systems and methods for actuating trastuzumab-conjugated liposomes accumulated in breast cancer tissues for the treatment of breast cancer.

## Claims

### Claim 1

A method of treating breast cancer in a mammal, the method comprising:
delivering an actively targeted liposome to the mammal, the actively targeted liposome comprising:
a lipid bilayer forming a spherical shell, wherein the spherical shell comprises an interior liposomal cavity, wherein the lipid bilayer comprises:
cholesterol,
dipalmitoylphosphatidyl choline (DPPC),
1,2-distearoyl-sn-glycero-3-phosphoethanolamine-N-[amino(polyethylene glycol)-2000](DSPE-PEG(2000)-NH2); and
a plurality of trastuzumab molecules linked to a surface of the actively targeted liposome, each liposome comprising about 1 to 18 trastuzumab, and
a chemotherapeutic drug, the chemotherapeutic drug comprising at least one of a hydrophilic chemotherapeutic drug contained within the interior liposomal cavity and a hydrophobic chemotherapeutic drug contained within the lipid bilayer of the actively targeted liposome; and
allowing the actively targeted liposome to circulate throughout a circulatory system of the mammal for a time sufficient to allow aggregation of a therapeutic quantity of actively targeted liposomes at a treatment area comprising a breast cancer; and
applying ultrasound to the treatment area such that the actively targeted liposome is critically disrupted thereby releasing the chemotherapeutic drug in the treatment area;
wherein the ultrasound applied to the treatment area comprises a low frequency ultrasound with a power density between about .002 W/ cm2 and .060 W/ cm2.

### Claim 2 (depends on 1)

The method of claim 1, wherein the ultrasound comprises a frequency of about 20 kHz.

### Claim 3 (depends on 1)

The method of claim 1, wherein the low frequency ultrasound applied to the treatment area is applied for less than about 6 minutes.

### Claim 4 (depends on 1)

The method of claim 1, wherein pulsed ultrasound is applied to the treatment area.

### Claim 5 (depends on 1)

The method of claim 1, wherein the lipid bilayer of the actively targeted liposome comprises one or more PEGylated lipids.

### Claim 6 (depends on 5)

The method of claim 5, wherein the plurality of trastuzumab molecules are linked to a distal end of the PEG chain.

### Claim 7 (depends on 1)

The method of claim 1, wherein the actively targeted liposome comprises a protein density of 7.5-30 molecules per liposome.

### Claim 8 (depends on 1)

The method of claim 1, wherein the actively targeted liposome comprises a mean radius between 50-150 nm.

### Claim 9 (depends on 1)

The method of claim 1, wherein the actively targeted liposome comprises 6 to 12 trastuzumab molecules.

### Claim 10 (depends on 4)

The method of claim 4, wherein the ultrasound is pulsed 2 times.

### Claim 11 (depends on 4)

The method of claim 4, wherein the ultrasound is pulsed 3 times.

### Claim 12 (depends on 1)

The method of claim 1, wherein the chemotherapeutic drug is selected from the group of doxorubicin, annamycin, daunorubicin, vincristine, cisplatin derivatives, paclitaxel, 5-fluorouracil derivatives, camptothecin derivatives, and retinoids.

### Claim 13 (depends on 1)

The method of claim 1, wherein the actively targeted liposome comprises a plurality of vesicles, the plurality of vesicles linked to about 9 trastuzumab molecules.

### Claim 14 (depends on 1)

The method of claim 1, wherein the plurality of trastuzumab molecules are linked to the surface of the actively targeted liposome using cyanuric chloride.

### Claim 15 (depends on 1)

The method of claim 1, wherein 20% or more of the chemotherapeutic drug is released within 20 seconds after applying ultrasound to the treatment area.

### Claim 16 (depends on 1)

The method of claim 1, wherein 80% or more of the chemotherapeutic drug is released within 300 seconds after applying ultrasound to the treatment area.

### Claim 17 (depends on 2)

The method of claim 2, wherein the low frequency ultrasound comprises a power density of one of 7.46 mW/cm2, 9.85 mW/cm2, and 17.31 mW/cm2.

### Claim 18 (depends on 7)

The method of claim 7, wherein the ratio of PEGylated lipids to non-PEGylated lipids is below 10%.

### Claim 19 (depends on 1)

The method of claim 1, wherein the molar ratio of cholesterol, DPPC, and (DSPE-PEG(2000)-NH2) is about 30:65:5.

### Claim 20 (depends on 12)

The method of claim 12, wherein the chemotherapeutic drug is doxorubicin.