US5000960A — Protein coupling to lipid vesicles

Claim 1

Independent Claim 121 words

A method of coupling targeting molecules to nonphospholipid lipid vesicles while retaining the targeting specificity of said targeting molecules comprising the steps of: selecting a targeting molecule having a free NH 2 group thereon, reacting said targeting molecule with a bifunctional agent comprising N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate, in a manner such that said bifunctional agent reacts with said NH 2 group on said targeting molecule to incorporate a free SH group onto said targeting molecule, forming a nonphospholipid lipid vesicle having a steroid with a free SH group as one of its structural components, and reacting said lipid vesicle having a free SH group thereon and said targeting molecule with said free SH group, thereby coupling said targeting molecule to said lipid vesicle.

Claim 2

Independent Claim 121 words

A method of coupling targeting molecules to nonphospholipid lipid vesicles while retaining the targeting specificity of said targeting molecules comprising the steps of: selecting a targeting molecule having a free NH 2 group thereon, reacting said targeting molecule with a bifunctional agent comprising N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate, in a manner such that said bifunctional agent reacts with said NH 2 group on said targeting molecule to incorporate a free SH group onto said targeting molecule, forming a nonphospholipid lipid vesicle having a steroid with a free SH group as one of its structural components, and reacting said lipid vesicle having a free SH group thereon and said targeting molecule with said free SH group, thereby coupling said targeting molecule to said lipid vesicle.

Claim 3

Dependent Claim 15 words

The method of claim 1 wherein said steroid with a free SH group comprises thiocholesterol.

Claim 4

Dependent Claim 44 words

The method of claim 1 wherein said nonphospholipid lipid vesicle comprises polyoxyethylene fatty esters, polyoxyethylene fatty acid ethers, diethanolamines, long-chain acyl amino acid amides, long-chain acyl amides, polyoxyethylene sorbitan mono and tristearates and oleates, polyoxyethylene glyceryl monostearates and monooleates, and glyceryl monostearates and monooleates.

Claim 5

Dependent Claim 15 words

The method of claim 3 wherein said nonphospholipid lipid vesicle comprises a paucilamellar lipid vesicle.

Claim 6

Dependent Claim 19 words

The method of claim 4 wherein said nonphospholipid lipid vesicle further comprises a charge-producing agent as a structural component.

Claim 7

Dependent Claim 18 words

The method of claim 1 wherein said targeting molecule is selected from a group consisting of peptide hormones.

Claim 8

Dependent Claim 12 words

The method of claim 1 wherein said targeting molecule comprises a protein.

Claim 9

Dependent Claim 18 words

The method of claim 7 wherein said protein is selected from a group consisting of lectins, and immunoglobulins.

Claim 10

Dependent Claim 21 words

The method of claim 8 wherein said lectin is selected from a group consisting of concanavalin A, and wheat germ agglutinin.

Claim 11

Dependent Claim 24 words

The method of claim 8 wherein said immunoglobulin is selected from a group consisting of monoclonal antibodies, chimeric antibodies, and portions and fragments thereof.

Claim 12

Independent Claim 177 words

A method of delivering a specified material to a particular location or tissue in a human or animal body comprising the steps of: encapsulating said specified material in a nonphospholipid lipid vesicle, said lipid vesicle containing a steroid having a free SH group as one of its structural components, selecting a proteinaceous targeting molecule which reacts with said particular location or tissue, reacting said proteinaceous targeting molecule with a bifunctional agent comprising N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate, in a manner such that said bifunctional agent reacts with free NH 2 groups on said targeting molecule and provides a free SH group, thereby forming a modified targeting molecule, reacting a plurality of said modified targeting molecules with said lipid vesicle so that at least one of said modified targeting molecules is bound to the surface of said lipid vesicle by an S--S bond, introducing said lipid vesicle having said modified targeting molecule bound thereto into said human or animal body, and allowing said targeting molecule to react with its target, thereby bringing said encapsulated material to said specified location or tissue.

Claim 13

Independent Claim 177 words

A method of delivering a specified material to a particular location or tissue in a human or animal body comprising the steps of: encapsulating said specified material in a nonphospholipid lipid vesicle, said lipid vesicle containing a steroid having a free SH group as one of its structural components, selecting a proteinaceous targeting molecule which reacts with said particular location or tissue, reacting said proteinaceous targeting molecule with a bifunctional agent comprising N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate, in a manner such that said bifunctional agent reacts with free NH 2 groups on said targeting molecule and provides a free SH group, thereby forming a modified targeting molecule, reacting a plurality of said modified targeting molecules with said lipid vesicle so that at least one of said modified targeting molecules is bound to the surface of said lipid vesicle by an S--S bond, introducing said lipid vesicle having said modified targeting molecule bound thereto into said human or animal body, and allowing said targeting molecule to react with its target, thereby bringing said encapsulated material to said specified location or tissue.

Claim 14

Dependent Claim 15 words

The method of claim 11 wherein said nonphospholipid lipid vesicle comprises a paucilamellar lipid vesicle.

Claim 15

Dependent Claim 18 words

The method of claim 11 wherein said proteinaceous targeting molecule is selected from a group consisting of immunoglobulins.

Claim 16

Dependent Claim 24 words

The method of claim 13 wherein said immunoglobulin is selected from a group consisting of monoclonal antibodies, chimeric antibodies, and fragments and portions thereof.

Claim 17

Dependent Claim 59 words

A nonphospholipid lipid vesicle having a targeting molecule attached thereto comprising a nonphospholipid lipid vesicle containing a steroid having free SH groups as one of its structural components, said targeting molecule being attached to said lipid vesicle by an S--S bond between a free SH group on said targeting molecule and a free SH group on said lipid vesicle.

Claim 18

Dependent Claim 59 words

A nonphospholipid lipid vesicle having a targeting molecule attached thereto comprising a nonphospholipid lipid vesicle containing a steroid having free SH groups as one of its structural components, said targeting molecule being attached to said lipid vesicle by an S--S bond between a free SH group on said targeting molecule and a free SH group on said lipid vesicle.

Claim 19

Dependent Claim 15 words

The lipid vesicle of claim 15 wherein said lipid vesicle comprises a paucilamellar lipid vesicle.

Claim 20

Dependent Claim 21 words

The lipid vesicle of claim 15 wherein said targeting molecule is selected from a group consisting of proteins, and peptide hormones.

Claim 21

Dependent Claim 24 words

The lipid vesicles of claim 17 targeting molecule is a protein selected from a group consisting of lectins, immunoglobulins, monoclonal antibodies, and chimeric antibodies.

Claim 22

Dependent Claim 49 words

The lipid vesicles of claim 15 wherein said targeting molecule is attached to said lipid vesicle by a bifunctional agent comprising N-hydroxysuccinimidyl 3-(2-pyridyldithio)propionate, in a manner such that said bifunctional agent attaches to a free NH 2 group on said targeting molecule and supplies a free SH group thereto.

Claim 23

Dependent Claim 15 words

The lipid vesicle of claim 15 wherein said vesicle has a diagnostic agent encapsulated therein.

Claim 24

Dependent Claim 13 words

The lipid vesicle of claim 20 wherein said targeting molecule comprises an immunoglobulin.

Claim 25

Dependent Claim 25 words

The lipid vesicle of claim 21 wherein said immunoglobulin is selected from a group consisting of monoclonal antibodies, chimeric antibodies, and portions and fragments thereof.

Claim 26

Dependent Claim 27 words

The lipid vesicle of claim 20 wherein said diagnostic agent is selected from a group consisting of immunodiagnostic agents, visualization agents, high density particles and magnetic particles.

Annuity Stage	Window Opens	Payment Deadline	Surcharge Period Until	Estimated Fee	Status
3.5 Years (Year 4)	1994-03-18	1994-09-17	1995-03-19	$2000.00	Window Closed / Lapsed
7.5 Years (Year 8)	1998-03-18	1998-09-17	1999-03-19	$3760.00	Window Closed / Lapsed
11.5 Years (Year 12)	2002-03-18	2002-09-17	2003-03-19	$7700.00	Window Closed / Lapsed

Protein coupling to lipid vesicles

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