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US-12622953-B2 - Vaccines targeting neoepitopes

US12622953B2US 12622953 B2US12622953 B2US 12622953B2US-12622953-B2

Abstract

Methods of vaccination utilizing a cationic liposomal adjuvant admixed with at least one neoepitope and a solvent. Also provided are unit dosages and compositions for use in the methods.

Inventors

  • Jens Kringelum
  • Anders Bundgård Sørensen
  • Birgitte Rønø
  • Nadia Viborg Petersen
  • Signe Tandrum Schmidt
  • Lars Vibe Andreasen
  • Dennis Christensen

Assignees

  • EVAXION BIOTECH A/S
  • Staten Serum Institut

Dates

Publication Date
20260512
Application Date
20200103
Priority Date
20190103

Claims (20)

  1. 1 . A unit dose of an immunogenic composition, said unit dose comprising an immunogenic effective amount of at least one peptide (non-peptide), which comprises an amino acid sequence of a neo-epitope of a patient's neoplastic cells, a cationic liposomal adjuvant, and a solvent, wherein the cationic liposomal adjuvant comprises dimethyldioctadecylammonium (DDA), polyinosinic acid:polycytidylic acid (poly(I:C)), and MMG, wherein MMG is monomycolyl glycerol or its synthetic analogue 3-hydroxy-2-tetradecyl-octadecanoic acid-2,3-dihydroxypropyl ester, and wherein the unit dose comprises at least 100 μg of each of said at least one neo-peptide.
  2. 2 . The unit dose according to claim 1 , which comprises at least 100 μg, at least or at most 105 μg, at least or at most 110 μg, at least or at most 115 μg, at least or at most 120 μg, at least or at most 125 μg, at least or at most 130 μg, at least or at most 135 μg, at least or at most 140 μg, at least or at most 145 μg, at least or at most 150 μg, at least or at most 155 μg, at least or at most 160 μg, at least or at most 165 μg, at least or at most 170 μg, at least or at most 175 μg, at least or at most 180 μg, at least or at most 185 μg, at least or at most 190 μg, at least or at most 195 μg, at least or at most 200 μg, at least or at most 205 μg, at least or at most 210 μg, at least or at most 215 μg, at least or at most 220 μg, at least or at most 225 μg, at least or at most 230 μg, at least or at most 235 μg, at least or at most 240 μg, at least or at most 245 μg, at least or at most 250 μg, at least or at most 255 μg, at least or at most 260 μg, at least or at most 265 μg, at least or at most 270 μg, at least or at most 275 μg, at least or at most 280 μg, at least or at most 285 μg, at least or at most 290 μg, at least or at most 295 μg, at least or at most 300 μg, at least or at most 305 μg, at least or at most 310 μg, at least or at most 315 μg, at least or at most 320 μg, at least or at most 325 μg, at least or at most 330 μg, at least or at most 335 μg, at least or at most 340 μg, at least or at most 345 μg, at least or at most 350 μg, at least or at most 355 μg, at least or at most 360 μg, at least or at most 365 μg, at least or at most 370 μg, at least or at most 375 μg, at least or at most 380 μg, at least or at most 385 μg, at least or at most 390 μg, at least or at most 395 μg, or at least or at most 400 μg of each neo-peptide.
  3. 3 . The unit dose according to claim 2 , which comprises an amount selected from 100 μg, 101 μg, 102 μg, 103 μg, 104 μg, 105 μg, 106 μg, 107 μg, 108 μg, 109 μg, 110 μg, 111 μg, 112 μg, 113 μg, 114 μg, 115 μg, 116 μg, 117 μg, 118 μg, 119 μg, 120 μg, 121 μg, 122 μg, 123 μg, 124 μg, 125 μg, 126 μg, 127 μg, 128 μg, 129 μg, 130 μg, 131 μg, 132 μg, 133 μg, 134 μg, 135 μg, 136 μg, 137 μg, 138 μg, 139 μg, 140 μg, 141 μg, 142 μg, 143 μg, 144 μg, 145 μg, 146 μg, 147 μg, 148 μg, 149 μg, 150 μg, 151 μg, 152 μg, 153 μg, 154 μg, 155 μg, 156 μg, 157 μg, 158 μg, 159 μg, 160 μg, 161 μg, 162 μg, 163 μg, 164 μg, 165 μg, 166 μg, 167 μg, 168 μg, 169 μg, 170 μg, 171 μg, 172 μg, 173 μg, 174 μg, 175 μg, 176 μg, 177 μg, 178 μg, 179 μg, 180 μg, 181 μg, 182 μg, 183 μg, 184 μg, 185 μg, 186 μg, 187 μg, 188 μg, 189 μg, 190 μg, 191 μg, 192 μg, 193 μg, 194 μg, 195 μg, 196 μg, 197 μg, 198 μg, 199 μg, 200 μg, 201 μg, 202 μg, 203 μg, 204 μg, 205 μg, 206 μg, 207 μg, 208 μg, 209 μg, 210 μg, 211 μg, 212 μg, 213 μg, 214 μg, 215 μg, 216 μg, 217 μg, 218 μg, 219 μg, 220 μg, 221 μg, 222 μg, 223 μg, 224 μg, 225 μg, 226 μg, 227 μg, 228 μg, 229 μg, 230 μg, 231 μg, 232 μg, 233 μg, 234 μg, 235 μg, 236 μg, 237 μg, 238 μg, 239 μg, 240 μg, 241 μg, 242 μg, 243 μg, 244 μg, 245 μg, 246 μg, 247 μg, 248 μg, 249 μg, 250 μg, 251 μg, 252 μg, 253 μg, 254 μg, 255 μg, 256 μg, 257 μg, 258 μg, 259 μg, 260 μg, 261 μg, 262 μg, 263 μg, 264 μg, 265 μg, 266 μg, 267 μg, 268 μg, 269 μg, 270 μg, 271 μg, 272 μg, 273 μg, 274 μg, 275 μg, 276 μg, 277 μg, 278 μg, 279 μg, 280 μg, 281 μg, 282 μg, 283 μg, 284 μg, 285 μg, 286 μg, 287 μg, 288 μg, 289 μg, 290 μg, 291 μg, 292 μg, 293 μg, 294 μg, 295 μg, 296 μg, 297 μg, 298 μg, 299 μg, 300 μg, 301 μg, 302 μg, 303 μg, 304 μg, 305 μg, 306 μg, 307 μg, 308 μg, 309 μg, 310 μg, 311 μg, 312 μg, 313 μg, 314 μg, 315 μg, 316 μg, 317 μg, 318 μg, 319 μg, 320 μg, 321 μg, 322 μg, 323 μg, 324 μg, 325 μg, 326 μg, 327 μg, 328 μg, 329 μg, 330 μg, 331 μg, 332 μg, 333 μg, 334 μg, 335 μg, 336 μg, 337 μg, 338 μg, 339 μg, 340 μg, 341 μg, 342 μg, 343 μg, 344 μg, 345 μg, 346 μg, 347 μg, 348 μg, 349 μg, 350 μg, 351 μg, 352 μg, 353 μg, 354 μg, 355 μg, 356 μg, 357 μg, 358 μg, 359 μg, 360 μg, 361 μg, 362 μg, 363 μg, 364 μg, 365 μg, 366 μg, 367 μg, 368 μg, 369 μg, 370 μg, 371 μg, 372 μg, 373 μg, 374 μg, 375 μg, 376 μg, 377 μg, 378 μg, 379 μg, 380 μg, 381 μg, 382 μg, 383 μg, 384 μg, 385 μg, 386 μg, 387 μg, 388 μg, 389 μg, 390 μg, 391 μg, 392 μg, 393 μg, 394 μg, 395 μg, 396 μg, 397 μg, 398 μg, 399 μg, and 400 μg of each neo-peptide.
  4. 4 . The unit dose according to claim 1 , wherein the number of neo-peptides is selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30.
  5. 5 . The unit dose according to claim 1 , wherein the ratio DDA:MMG is 5:1 by (w/w), and wherein the ratio DDA:poly(I:C) is between 5:1 and 20:1 (w/w).
  6. 6 . The unit dose according to claim 1 , wherein the DDA:MMG:poly(I:C) relative weights are 5:1:1.
  7. 7 . The unit dose according to claim 1 , wherein the DDA:MMG:poly(I:C) relative weights are 20:4:1.
  8. 8 . The unit dose according to claim 1 , wherein the at least one neo-peptide is free from cysteine residues.
  9. 9 . The unit dose according to claim 1 , wherein the at least one neo-peptide is free from Q and N as the N-terminal amino acid residue.
  10. 10 . The unit dose according to claim 1 , wherein the at least one neo-peptide is free from the amino acid sequence DG in the N-terminus.
  11. 11 . The unit dose according to claim 1 , wherein the immunogenic composition has a pH in the range between 7.0 and 7.6.
  12. 12 . The unit dose according to claim 11 , wherein the pH is about 7.4.
  13. 13 . A method for treatment of a neoplasm, in a mammalian patient, wherein the neoplasm exhibits T-cell epitopes (neo-epitopes) that are not exhibited by non-neoplastic cells in the patient, the method comprising administering an immunogenically effective amount of a liposomal composition comprising 1) at least one peptide (neo-peptide), which comprises an amino acid sequence of a neo-epitope of the patient's neoplastic cells, 2) a solvent, and 3) a cationic liposomal adjuvant wherein the cationic liposomal adjuvant comprises dimethyldioctadecylammonium (DDA), polyinosinic acid:polycytidylic acid (poly(I:C)), and MMG, wherein MMG is monomycolyl glycerol or its synthetic analogue 3-hydroxoy-2-tetradecyl-octadecanoic acid-2,3-dihydroxypropyl ester, and wherein the amount of each of the neo-peptide(s) administered is at least 100 μg.
  14. 14 . The method according to claim 13 , wherein the ratio DDA:MMG is 5:1 by (w/w), and wherein the ratio DDA:poly(I:C) is between 5:1 and 20:1 (w/w).
  15. 15 . The method according to claim 13 , wherein the DDA:MMG:poly(I:C) relative weights are 5:1:1.
  16. 16 . The method according to claim 13 , wherein the DDA:MMG:poly(I:C) relative weights are 20:4:1.
  17. 17 . The method according co claim 13 , wherein the at least one neo-peptide is water-soluble wherein 1.0 mg/ml neo-peptide in 25 mM TRIS pH 7.4 containing 5% (v/v) DMSO provides for an NTU (nephelometric turbidity unit) of at most 50, and 1.0 mg/ml neo-peptide in 25 mM TRIS pH 7.4 and 0.5% (v/v) DMSO provides for an NTU of at most 25.
  18. 18 . The method according to claim 13 , wherein the at least one neo-peptide exhibits at most 50% reduction in its concentration when in a mixture of a plurality subjected filtration to sterile when present in a mixture of a plurality of neo-peptides and 25 mM TRIS pH 7.4 containing 5% (v/v) DMSO, wherein each of the plurality of peptides have a concentration of 0.1 mg/ml in the mixture prior to sterile filtration.
  19. 19 . The method according to claim 13 , wherein the number of neo-peptides is selected from 2, 3, 4, 5, 6, 7, 8, 9, 10, 11, 12, 13, 14, 15, 16, 17, 18, 19, 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, and 30 neo-peptides.
  20. 20 . The method according to claim 13 , wherein the at least one neo-peptide includes a neo-epitope, which exhibits all MHC binding stability, which is above average, among neo-epitopes identified in the neoplastic cells.

Description

CROSS REFERENCE TO RELATED APPLICATIONS This is a national stage filing in accordance with 35 U.S.C. § 371 of PCT/EP2020/050058, filed Jan. 3, 2020, which claims the benefit of the priority of European Patent Application No. 19207238.7, filed Nov. 5, 2019 and European Patent Application No. 19150200.4, filed Jan. 3, 2019, the contents of each are incorporated herein by reference. FIELD OF THE INVENTION The present invention relates to the field of immunotherapy. In particular, the present invention relates to therapeutic immunization technologies for treatment of neoplastic diseases. BACKGROUND OF THE INVENTION Treatment of malignant neoplasms in patients has traditionally focussed on eradication/removal of the malignant tissue via surgery, radiotherapy, and/or chemotherapy using cytotoxic drugs in dosage regimens that aim at preferential killing of malignant cells over killing of non-malignant cells. In addition to the use of cytotoxic drugs, more recent approaches have focussed on targeting of specific biologic markers in the cancer cells in order to reduce systemic adverse effects exerted by classical chemotherapy. Monoclonal antibody therapy targeting cancer associated antigens has proven quite effective in prolonging life expectance in a number of malignancies. While being successful drugs, monoclonal antibodies that target cancer associated antigens or antigen can by their nature only be developed to target expression products that are known and appear in a plurality of patients, meaning that the vast majority of cancer specific antigens cannot be addressed by this type of therapy, because a large number of cancer specific antigens only appear in tumours from one single patient, cf. below. As early as in the late 1950'ies the theory of immunosurveillance proposed by Burnet and Thomas suggested that lymphocytes recognizes and eliminates autologous cells—including cancer cells—that exhibit altered antigenic determinants, and it is today generally accepted that the immune system inhibits carcinogenesis to a high degree. Nevertheless, immunosurveillance is not 100% effective and it is a continuing task to device cancer therapies where the immune system's ability to eradicate cancer cells is sought improved/stimulated. One approach has been to induce immunity against cancer-associated antigens, but even though this approach has the potential of being promising, it suffers the same drawback as antibody therapy that only a limited number of antigens can be addressed. Many if not all tumours express mutations. These mutations potentially create new targetable antigens (neo-antigens), which are potentially useful in specific T cell immunotherapy if it is possible to identify the neo-antigens and their antigenic determinants within a clinically relevant time frame. Since it with current technology is possible to fully sequence the genome of cells and to analyse for existence of altered or new expression products, it is possible to design personalized vaccines based neo-antigens. However, attempts at providing satisfactory clinical end-points have as today failed. There is hence an existing need for provision of anti-cancer vaccines that can effectively target neo-antigens and induce clinically significant immune responses in the vaccinated individuals. OBJECT OF THE INVENTION It is an object of embodiments of the invention to provide methods for induction of therapeutically effective immune responses against neo-antigens that comprise neo-epitopes. It is a further object to provide compositions comprising neo-epitope material that can be used for cancer immunotherapy. SUMMARY OF THE INVENTION It has been found by the present inventor(s) that formulation of neo-peptides with certain cationic liposomal adjuvants and administration of such formulations using a relatively high dosage of the neo-peptides provide for improved immune responses in vaccinated individuals. In turn, this is believed to provide for an improvement in cancer immunotherapy of cancers characterized by expression of neo-epitope containing expression products. So, in a first aspect the present invention relates to a method for treatment of a neoplasm, such as a malignant neoplasm, in a mammalian patient, wherein the neoplasm exhibits T-cell epitopes (neo-epitopes) that are not exhibited by non-neoplastic cells in the patient, the method comprising administering an immunogenically effective amount of a liposomal composition comprising 1) at least one peptide (neo-peptide), which comprises an amino acid sequence of a neo-epitope of the patient's neoplastic cells,2) a solvent, and3) a cationic liposomal adjuvant. A second aspect of the invention relates to a unit dose of an immunogenic composition, said unit dose comprising an immunogenically effective amount of at least one peptide (neo-peptide), which comprises an amino acid sequence of a neo-epitope of a patient's neoplastic cells, a cationic liposomal adjuvant, and a solvent. A third aspect of