# US 2014/0154282 A1 · Conjugates utilizing platform technology for stimulating immune response > Agents: this data is available as JSON from the patentgrep API and MCP server — see https://patentgrep.com/llms.txt - Number: US 2014/0154282 A1 - Document ID: US20140154282A1 - Status: Published application - Kind: A1 - Application number: 13/981,300 - Filed: 2012-01-26 - Published: 2014-06-05 - Earliest claimed: 2011-01-26 - Inventors: Robert S. Hodges; Kathryn V. Holmes; Zhe Yan; Wendy Jeanne Hartsock; Zhaohui Qian; Brooke Elizabeth Bishop Hirsch - Web page: https://patentgrep.com/patent/US20140154282A1 ## CPC - A61K 39/145: Orthomyxoviridae, e.g. influenza virus - A61K 39/12: Viral antigens - A61K 47/643: {Albumins, e.g. HSA, BSA, ovalbumin or a Keyhole Limpet Hemocyanin [KHL]} - A61P 31/16: for influenza or rhinoviruses - A61P 37/04: Immunostimulants - A61K 2039/6081: {Albumin; Keyhole limpet haemocyanin [KLH]} - A61K 2039/64: {characterised by the architecture of the carrier-antigen complex, e.g. repetition of carrier-antigen units} - C12N 2760/16134: Use of virus or viral component as vaccine, e.g. live-attenuated or inactivated virus, VLP, viral protein - 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) - A61P: SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS - C12N: MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA (microbiological testing media C12Q1/00) ## Abstract Templated conjugates created from naturally-occurring protein sequences found in pathogens, such as viruses, are disclosed. The sequences are “templated” into a consensus coiled-coil sequence in a platform in order to form a two-stranded antigen suitable for immunization of a subject. ## Claims ### Claim 1 A conjugate, comprising: two templated alpha helical polypeptides of approximately equal length, wherein each polypeptide comprises at least two heptad repeats, and wherein the polypeptides are derived from at least one virus; a covalent linkage between the two polypeptides; and a carrier protein covalently linked to one of the polypeptides; wherein the first polypeptide comprises the form: [I-b1i-c1i-L-e1i-f1i-g1i]n, where [I-b1i-c1i-L-e1i-f1i-g1i] is a pattern that repeats n times in the sequence of the first polypeptide, giving rise to at least two discrete seven-amino-acid segments, where I in each segment is isoleucine, L in each segment is leucine, n is an integer of at least 2; i is an integer from 1 to n, wherein the value of i is determined by the position of the segment in which it appears, such that the N-terminal segment which appears first in the sequence is assigned a value of i=1, i is incremented by one for each additional segment, and the C-terminal segment is assigned a value of i=n; where each b, c, e, f, and g in each of the n segments is selected independently of each b, c, e, f, and g amino acid in all other segments of the first polypeptide, and of all segments of the second polypeptide; the b, c, e, f, and g amino acids are selected from an epitope; the second polypeptide comprises the form: [I-b2i-c2i-L-e2i-f2i-g2i]n, where [I-b2i-c2i-L-e2i-f2i-g2i] is a pattern that repeats n times in the sequence of the second polypeptide, giving rise to at least two discrete seven-amino-acid segments, where I in each segment is isoleucine, L in each segment is leucine, n is an integer of at least 2 and is the same as n for the first polypeptide; i is an integer from 1 to n, wherein the value of i is determined by the position of the segment in which it appears, such that the N-terminal segment which appears first in the sequence is assigned a value of i=1, i is incremented by one for each additional segment, and the C-terminal segment is assigned a value of i=n; where each b, c, e, f, and g in each of the n segments is selected independently of each b, c, e, f, and g amino acid in all other segments of the second polypeptide, and of all segments of the first polypeptide; the b, c, e, f, and g amino acids are selected from an epitope which is different from the epitope of the first polypeptide; wherein the conjugate has the form: [Carrier Moiety]-[Linker A]-[Linker B1]-[Templated Epitope 1]-[Epitope 1 Modifier] [Modifier B2]-[Templated Epitope 2]-[Epitope 2 Modifier] where Carrier Moiety, Linker A, Linker B 1, Modifier B2, Epitope 1 Modifier, and Epitope 2 Modifier are optionally present; and optionally comprising an additional covalent Linker C between Templated Epitope 1 and Templated Epitope 2; optionally comprising an additional covalent Linker D between Epitope 1 Modifier and Epitope 2 Modifier, or optionally comprising an additional covalent Linker C between Templated Epitope 1 and Templated Epitope 2 and an additional covalent Linker D between Epitope 1 Modifier and Epitope 2 Modifier, wherein the Epitope 1 Modifier and the Epitope 2 Modifier are present and are selected from hydrophilic, polar, and charged amino acids, wherein the [Linker A] moiety is present, wherein the [Linker B1] moiety is present; with the proviso that either both Epitope 1 Modifier and Epitope 2 Modifier are present, or Linker C is present, or Linker D is present. ### Claim 2 (depends on 1) The conjugate of claim 1, wherein Templated Epitope 1 and Templated Epitope 2 are derived from two different epitope sequences chosen from the same strain of the same virus. ### Claim 3 (depends on 1) The conjugate of claim 1, wherein the epitope sequence from which Templated Epitope 1 is derived is chosen from a strain of a virus, and the epitope sequence from which Templated Epitope 2 is chosen is from the same epitope in a different strain of the same virus. ### Claim 4 (depends on 1) The conjugate of claim 1, wherein the epitope sequence from which Templated Epitope 1 is derived is chosen from a strain of a virus, and the epitope sequence from which Templated Epitope 2 is chosen is from a different epitope in a different strain of the same virus. ### Claim 5 (depends on 1) The conjugate of claim 1, wherein the epitope sequence from which Templated Epitope 1 is derived is chosen from a virus, and the epitope sequence from which Templated Epitope 2 is chosen is from a different virus. ### Claim 6 (depends on 1) The conjugate of claim 1, wherein the virus is an influenza virus. ### Claim 7 (depends on 5) The conjugate of claim 5, wherein one of Templated Epitope 1 and Templated Epitope 2 is derived from a sequence chosen from an influenza virus, and the other of Templated Epitope 1 and Templated Epitope 2 is derived from a sequence chosen from a virus other than influenza virus. ### Claim 8 (depends on 2) The conjugate of claim 2, wherein Templated Epitope 1 is Influenza PR8HA2 5P(420-448) Templated Epitope 5P (IENLNKKIDDLFLDIWTLNAEILVLENCRR-amide (SEQ ID NO: )) and Templated Epitope 2 is Influenza PR8HA2 6P(448-476) Templated Epitope 6P (IRTLDFHISNLKNLIEKLKSQIKNLAKECRR-amide (SEQ ID NO: )). ### Claim 9 (depends on 8) The conjugate of claim 8 of the form: wherein KLH is keyhole limpet hemocyanin, Nle is norleucine, and the vertical bar | between each C residue in each strand indicates a cystine disulfide bond. ### Claim 10 (depends on 8) The conjugate of claim 8 of the form: wherein KLH is keyhole limpet hemocyanin, Nle is norleucine, and the vertical bar | between each C residue in each strand indicates a cystine disulfide bond. ### Claim 11 (depends on 1) The conjugate of claim 1, wherein the carrier moiety is selected from a protein, keyhole limpet hemocyanin (KLH), bovine serum albumin (BSA), ovalbumin, tetanus toxoid, cholera subunit B, protein D from H. influenza, diphtheria toxoid, a promiscuous T-cell peptide epitope, a promiscuous human measles T cell peptide epitope, the peptide KLLSLIKGVIVHRLEGVE (SEQ ID NO: ), a non-proteinaceous moiety, a polysaccharide, or alginic acid (alginate). ### Claim 12 (depends on 1) The conjugate of claim 1, wherein the linkage between the carrier moiety and Linker A (if present), Linker B1 (if present in the absence of Linker A), or Templated Epitope 1 (if Linker A and Linker B1 are absent) is chemically definite. ### Claim 13 (depends on 1) A method of generating a protective immune response in a subject in need thereof, comprising administering the conjugate of claim 1 to a subject in a sufficient amount to produce the protective immune response. ### Claim 14 (depends on 1) A method of inducing an antibody response in an individual in need thereof, the method comprising administering the conjugate of claim 1 to an individual in need thereof in an amount sufficient to induce an antibody response in the individual. ### Claim 15 (depends on 14) The method of claim 14 wherein the antibody response is the production of a neutralizing antibody. ### Claim 16 (depends on 1) A method of generating a protective immune response in a subject in need thereof, comprising administering the conjugate of claim 1 to a subject in a sufficient amount to produce the protective immune response. ### Claim 17 (depends on 1) A method of inducing an antibody response in an individual in need thereof, the method comprising administering the conjugate of claim 1 to an individual in need thereof in an amount sufficient to induce an antibody response in the individual. ### Claim 18 (depends on 17) The method of claim 17 wherein the antibody response is the production of a neutralizing antibody.