Methods and compositions related to peptides and proteins with C-terminal elements

• US 11,260,133 B2
• Filed: 02/20/2009
• Issued: 03/01/2022
• Est. Priority Date: 02/21/2008
• Status: Active Grant

• Current Assignee: Sanford-Burnham Medical Research Institute (Sanford Burnham Prebys Medical Discovery Institute)
• Original Assignee: Sanford-Burnham Medical Research Institute (Sanford Burnham Prebys Medical Discovery Institute)
• Inventors: Ruoslahti, Erkki, Teesalu, Tambet, Sugahara, Kazuki
• Primary Examiner(s): Alstrum-Acevedo, James H
• Assistant Examiner(s): Martinez, Tara L
• Application Number:
  US12/390,061
• Publication Number:
  US 20090226372A1
• Time in Patent Office: 4,757 Days
• Field of Search: 530326
• CPC Class Codes: A61K 47/62 the modifying agent being a protein, peptide or polyamino acid
• CPC Class Codes: A61K 47/64 Drug-peptide, drug-protein or drug-polyamino acid conjugates, i.e. the modifying agent being a peptide, protein or polyamino acid which is covalently bonded or complexed to a therapeutically active agent peptidic linkers A61K47/65
• CPC Class Codes: A61K 47/6923 the form being an inorganic particle, e.g. ceramic particles, silica particles, ferrite or synsorb
• CPC Class Codes: A61P 35/00 Antineoplastic agents

Abstract

Disclosed are compositions and methods useful for targeting and internalizing molecules into cells of interest and for penetration by molecules of tissues of interest. The compositions and methods are based on peptide sequences that are selectively internalized by a cell, penetrate tissue, or both. The disclosed internalization and tissue penetration is useful for delivering therapeutic and detectable agents to cells and tissues of interest.

Claims

1. A method of forming a CendR conjugate, the method comprising

(a) selecting an amino acid sequence comprising a CendR element for internalization into a cell, tissue penetration, or both, wherein the CendR element is at a C-terminal end of the conjugate and has an exposed C-terminal carboxyl group, wherein the CendR element has the sequence X₁X₂X₃X₄, where X₁ can be R, or K, where X₄ can be R, K, or KG, and where X₂ and X₃ can each be, independently, any amino acid, wherein X₁ and X₄ are not both R, and wherein the CendR element does not include KLRK (SEQ ID NO:

168), KPPR (SEQ ID NO;

169), KRSR (SEQ ID NO;

170), KWKK (SEQ ID NO;

173), RRLK (SEQ ID NO;

178), RRTK (SEQ ID NO;

183), RRPK (SEQ ID NO;

184), KQRR (SEQ ID NO;

185), KRAR (SEQ ID NO;

186), KRGR (SEQ ID NO;

187), RSFK (SEQ ID NO;

188), KKPR (SEQ ID NO;

189), RLAK (SEQ ID NO;

174), or KRTR (SEQ ID NO;

191), and

(b) causing a cargo composition to be covalently coupled or non-covalently associated with a protein or peptide comprising the selected amino acid sequence,

wherein the cargo composition is coupled or associated with the protein or peptide on the N terminal side of the CendR element,

wherein the CendR conjugate comprises the protein or peptide and the coupled or associated cargo composition,

wherein the CendR element binds to neuropilin 1 (NRP-1), wherein internalization of the CendR element into cells is dependent on NRP-1, and

wherein the CendR element has the sequence KLAK (amino acids 11-14 of SEQ ID NO;

5), KPAR (amino acids 4-7 of SEQ ID NO;

114), KPER (amino acids 4-7 of SEQ ID NO;

90), KPLR (amino acids 4-7 of SEQ ID NO;

63), KPRR (amino acids 5-8 of SEQ ID NO;

126), KQAR (amino acids 4-7 of SEQ ID NO;

64), KRDR (amino acids 3-6 of SEQ ID NO;

89), KRER (amino acids 4-7 of SEQ ID NO;

91), KTAR (amino acids 3-6 of SEQ ID NO;

56), RGDK (amino acids 2-5 of SEQ ID NO;

4), RPAK (amino acids 4-7 of SEQ ID NO;

112), or RTPK (amino acids 4-7 of SEQ ID NO;

84).

2. A CendR conjugate made by the method comprising

(a) selecting an amino acid sequence for internalization into a cell, tissue penetration, or both, wherein the amino acid sequence comprises a CendR element, wherein the CendR element is at a C-terminal end of the conjugate and has an exposed C-terminal carboxyl group, wherein the CendR element has the sequence X₁X₂X₃X₄, where X₁ can be R, or K, where X₄ can be R, K, or KG, and where X₂ and X₃ can each be, independently, any amino acid, wherein X₁ and X₄ are not both R, and wherein the CendR element does not include KLRK (SEQ ID NO:

168), KPPR (SEQ ID NO;

169), KRSR (SEQ ID NO;

170), KWKK (SEQ ID NO;

173), RRLK (SEQ ID NO;

178), RRTK (SEQ ID NO;

183), RRPK (SEQ ID NO;

184), KQRR (SEQ ID NO;

185), KRAR (SEQ ID NO;

186), KRGR (SEQ ID NO;

187), RSFK (SEQ ID NO;

188), KKPR (SEQ ID NO;

189), RLAK (SEQ ID NO;

174), or KRTR (SEQ ID NO;

191), and

(b) causing a cargo composition to be covalently coupled or non-covalently associated with a protein or peptide comprising the selected amino acid sequence,

wherein the cargo composition is coupled or associated with the protein or peptide on the N terminal side of the CendR element,

wherein the CendR conjugate comprises the protein or peptide and the coupled or associated cargo composition,

wherein the CendR element binds to neuropilin 1 (NRP-1), wherein internalization of the CendR element into cells is dependent on NRP-1, and

wherein the CendR element has the sequence KLAK (amino acids 11-14 of SEQ ID NO;

5), KPAR (amino acids 4-7 of SEQ ID NO;

114), KPER (amino acids 4-7 of SEQ ID NO;

90), KPLR (amino acids 4-7 of SEQ ID NO;

63), KPRR (amino acids 5-8 of SEQ ID NO;

126), KQAR (amino acids 4-7 of SEQ ID NO;

64), KRDR (amino acids 3-6 of SEQ ID NO;

89), KRER (amino acids 4-7 of SEQ ID NO;

91), KTAR (amino acids 3-6 of SEQ ID NO;

56), RGDK (amino acids 2-5 of SEQ ID NO;

4), RPAK (amino acids 4-7 of SEQ ID NO;

112), or RTPK (amino acids 4-7 of SEQ ID NO;

84).