DETAILED ACTION
Notice of Pre-AIA or AIA Status
The present application, filed on or after March 16, 2013, is being examined under the first inventor to file provisions of the AIA .
1. Claims 1-3, 7-15, 17-23 are pending and being examined.
Claim Objections
2. Claim 10 is objected to because of the following informalities: Claim 10 appears to have a typo reciting: “wherein the FAF1 of SEQ ID NO:1 is loaded in the exosomes.” It appears the claim should be corrected to recite: “wherein the FAF1 protein comprising SEQ ID NO:1 is loaded in the exosomes.”
Claim 10 is also objected to under 37 CFR 1.75(c), as being of improper dependent form for failing to further limit the subject matter of a previous claim. Applicant is required to cancel the claim(s), or amend the claim(s) to place the claim(s) in proper dependent form, or rewrite the claim(s) in independent form. Claim 1 requires FAF1 protein to be loaded in exosomes and claim 2 recites FAF1 comprises SEQ ID NO:1, therefore claim 10 depending from claim 2 and reciting that the FAF1 protein comprising SEQ ID NO:1 is loaded in the exosome, fails to further limit the claims.
3. Claim 15 is objected to because of the following informalities: Claim 15 recites a typo: “wherein the subject has having decreased expression of FAF1”. It appears the claim should recite: “wherein the subject has decreased expression of FAF1”. Appropriate correction is required.
4. Claim 23 is objected to because of the following informalities: Claim 23 recites a typo: “The method of claim 15, which further administering an additional therapeutic agent”. It appears the claim should recite: “The method of claim 15, further comprising administering to the subject an additional therapeutic agent”. Appropriate correction is required.
Claim Rejections - 35 USC § 112
The following is a quotation of 35 U.S.C. 112(b):
(b) CONCLUSION.—The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the inventor or a joint inventor regards as the invention.
The following is a quotation of 35 U.S.C. 112 (pre-AIA ), second paragraph:
The specification shall conclude with one or more claims particularly pointing out and distinctly claiming the subject matter which the applicant regards as his invention.
5. Claim 7 is rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. Claim 7 recites: “wherein the exosomes of the composition deliver FAF1 to target tumor cells.” Claim 7 depends from claim 1 composition/product claim, and it is unclear if the language in claim 7 is reciting a method step of delivering FAF1 to target cells and is drawn to a process invention, or if it is reciting a functional characteristic of the exosome and is drawn to a product invention. The metes and bounds of the claim cannot be determined. Clarification is required.
Claim 7 interpretation: For the sake of compact prosecution, Examiner is currently interpreting “deliver FAF1 to target tumors cells” as a functional feature of the exosome and not as a delivery method step.
6. Claims 8 and 9 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention.
Claim 8 depends from product claim 1, however, claim 8 recites a method step of “the composition is administered”.
Claim 9 depends from product claim 1, however, claim 9 recites a method step of “the composition is administered alone or concurrently and sequentially with other therapeutic agents.”
Claims 8 and 9 are unclear whether they are drawn to a product or a process invention. The metes and bounds of the claims cannot be determined.
Claim interpretation for claims 8 and 9: For the sake of compact prosecution, Examiner is currently interpreting “is administered” in claim 8 and 9 as an intended use of the claimed composition that does not alter the structure or function of the composition of claim 1.
7. Claims 13 and 14 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. Claim 13 recites: “1) treating, with a test substance, cancer cells having decreased expression of FAF1”. The claim is unclear with regard to what amount of FAF1 is “decreased” and scope of cells are encompassed by this, including in the cells listed in claim 14. What amount of FAF1 constitutes “decreased”? The metes and bounds of the claims cannot be determined and the scope of cells to be used in the claimed method cannot be determined.
8. Claims 15 and 17-23 are rejected under 35 U.S.C. 112(b) or 35 U.S.C. 112 (pre-AIA ), second paragraph, as being indefinite for failing to particularly point out and distinctly claim the subject matter which the inventor or a joint inventor (or for applications subject to pre-AIA 35 U.S.C. 112, the applicant), regards as the invention. Claim 15 recites: “wherein the subject has having decreased expression of FAF1”. The claim is unclear with regard to what amount or expression level of FAF1 is “decreased” and what subjects are encompassed by this. What level of FAF1 constitutes “decreased”? The metes and bounds of the claims cannot be determined and the scope of subjects treated by the claimed method cannot be determined.
Claim Rejections - 35 USC § 101
35 U.S.C. 101 reads as follows:
Whoever invents or discovers any new and useful process, machine, manufacture, or composition of matter, or any new and useful improvement thereof, may obtain a patent therefor, subject to the conditions and requirements of this title.
9. Claims 1-3, 7-10 are rejected under 35 U.S.C. 101 because the claimed invention is directed to a product of nature without significantly more. The claim(s) recite(s) a pharmaceutical composition comprising FAF1 protein-loaded exosomes, wherein the protein comprises SEQ ID NO:1, and wherein the exosomes have a diameter of 50-200 nm. The claimed composition is a product of nature. For example, WO 2021/193802, Ueno, discloses human patient-derived plasma (a pharmaceutical composition) containing exosomes identified as containing Fas-associated factor 1 (FAF1) protein encoded by NCBI accession number NP_008982 (see Example 1; Table 1-1 and Table 4-4, Table 5-4), which is 100% identical to instant SEQ ID NO:1 (see sequence alignment below). Carvalho et al (Frontiers in Bioscience, Landmark, Jan 1, 2020; 25:398-436) teach FAF1 protein-loaded exosomes (extracellular vesicles, EVs) are produced naturally in cancer cells and at a size of 30-120 nm (Figure 1; section 5; Table 1 melanoma cell lines). Hurwitz et al (Oncotarget, 2016, 7(No. 52):86999-87015) teaches naturally occurring exosomes isolated from breast cancer cells are loaded with FAF1 protein (see prior art rejection below).
This judicial exception is not integrated into a practical application because there is no method recited in the rejected claims employing the composition. The claim(s) does/do not include additional elements that are sufficient to amount to significantly more than the judicial exception because the claimed composition is not remarkably different from what occurs in nature.
Claim 7 fails to recite any additional structures or functions that include additional elements that are sufficient to amount to significantly more than the judicial exception and fails to distinguish the composition from what occurs in nature.
Claims 8 and 9 are unclear if a method of practical application is being claimed for the reasons stated in 35 U.S.C. 112(b) above, and the claims can be interpreted as reciting an intended use of the composition.
The claims fail to integrate the natural product into a practical application and fail to include additional elements that are sufficient to amount to significantly more than the judicial exception because the claimed composition is not remarkably different from what occurs in nature.
Instant SEQ ID NO:1 (Qy) aligned with NCBI NP_008982 FAS-associated factor 1 [Homo sapiens] (Db):
RESULT 1
AASEQ2_01302026_171529
Query Match 100.0%; Score 3342; DB 1; Length 650;
Best Local Similarity 100.0%;
Matches 650; Conservative 0; Mismatches 0; Indels 0; Gaps 0;
Qy 1 MASNMDREMILADFQACTGIENIDEAITLLEQNNWDLVAAINGVIPQENGILQSEYGGET 60
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 1 MASNMDREMILADFQACTGIENIDEAITLLEQNNWDLVAAINGVIPQENGILQSEYGGET 60
Qy 61 IPGPAFNPASHPASAPTSSSSSAFRPVMPSRQIVERQPRMLDFRVEYRDRNVDVVLEDTC120 ----- ||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 61 IPGPAFNPASHPASAPTSSSSSAFRPVMPSRQIVERQPRMLDFRVEYRDRNVDVVLEDTC120
Qy 121 TVGEIKQILENELQIPVSKMLLKGWKTGDVEDSTVLKSLHLPKNNSLYVLTPDLPPPSSS180
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 121 TVGEIKQILENELQIPVSKMLLKGWKTGDVEDSTVLKSLHLPKNNSLYVLTPDLPPPSSS180
Qy 181 SHAGALQESLNQNFMLIITHREVQREYNLNFSGSSTIQEVKRNVYDLTSIPVRHQLWEGW240
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 181 SHAGALQESLNQNFMLIITHREVQREYNLNFSGSSTIQEVKRNVYDLTSIPVRHQLWEGW240
Qy 241 PTSATDDSMCLAESGLSYPCHRLTVGRRSSPAQTREQSEEQITDVHMVSDSDGDDFEDAT300
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 241 PTSATDDSMCLAESGLSYPCHRLTVGRRSSPAQTREQSEEQITDVHMVSDSDGDDFEDAT300
Qy 301 EFGVDDGEVFGMASSALRKSPMMPENAENEGDALLQFTAEFSSRYGDCHPVFFIGSLEAA360
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 301 EFGVDDGEVFGMASSALRKSPMMPENAENEGDALLQFTAEFSSRYGDCHPVFFIGSLEAA360
Qy 361 FQEAFYVKARDRKLLAIYLHHDESVLTNVFCSQMLCAESIVSYLSQNFITWAWDLTKDSN420
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 361 FQEAFYVKARDRKLLAIYLHHDESVLTNVFCSQMLCAESIVSYLSQNFITWAWDLTKDSN420
Qy 421 RARFLTMCNRHFGSVVAQTIRTQKTDQFPLFLIIMGKRSSNEVLNVIQGNTTVDELMMRL480
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 421 RARFLTMCNRHFGSVVAQTIRTQKTDQFPLFLIIMGKRSSNEVLNVIQGNTTVDELMMRL480
Qy 481 MAAMEIFTAQQQEDIKDEDEREARENVKREQDEAYRLSLEADRAKREAHEREMAEQFRLE540
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 481 MAAMEIFTAQQQEDIKDEDEREARENVKREQDEAYRLSLEADRAKREAHEREMAEQFRLE540
Qy 541 QIRKEQEEEREAIRLSLEQALPPEPKEENAEPVSKLRIRTPSGEFLERRFLASNKLQIVF600
||||||||||||||||||||||||||||||||||||||||||||||||||||||||||||
Db 541 QIRKEQEEEREAIRLSLEQALPPEPKEENAEPVSKLRIRTPSGEFLERRFLASNKLQIVF600
Qy 601 DFVASKGFPWDEYKLLSTFPRRDVTQLDPNKSLLEVKLFPQETLFLEAKE 650
||||||||||||||||||||||||||||||||||||||||||||||||||
Db 601 DFVASKGFPWDEYKLLSTFPRRDVTQLDPNKSLLEVKLFPQETLFLEAKE 650
Claim Rejections - 35 USC § 102
The following is a quotation of the appropriate paragraphs of 35 U.S.C. 102 that form the basis for the rejections under this section made in this Office action:
A person shall be entitled to a patent unless –
(a)(1) the claimed invention was patented, described in a printed publication, or in public use, on sale, or otherwise available to the public before the effective filing date of the claimed invention.
(a)(2) the claimed invention was described in a patent issued under section 151, or in an application for patent published or deemed published under section 122(b), in which the patent or application, as the case may be, names another inventor and was effectively filed before the effective filing date of the claimed invention.
Claim interpretation for claims 8 and 9: For the sake of compact prosecution, Examiner is currently interpreting “is administered” in claim 8 and 9 as an intended use of the claimed composition that does not alter the structure or function of the composition of claim 1.
10. Claim(s) 1, 3, 7-9 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Hurwitz et al (Oncotarget, 2016, 7(No. 52):86999-87015).
Hurwitz teaches isolating exosomes (extracellular vesicles, EVs) in the range of 40-150 nm diameter from cancer cell lines, wherein the exosomes are comprised in PBS solution (pharmaceutical composition) (Introduction p. 86999, col. 1; Figure 1A; “EV enrichment and protein quantification” p. 87009); and wherein the exosomes are loaded with FAF1 protein (Supplementary Table S2, see the excerpt below copied and pasted from the supplementary Excel file of Table S2 demonstrating FAF1 protein-loaded exosomes were isolated from breast cancer MDA-MB-468 cells at item #5804):
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60
900
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21
908
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Greyscale
11. Claim(s) 1, 2, 7-10 are rejected under 35 U.S.C. 102(a)(2) as being anticipated by WO 2021/193802, Ueno et al, filed March 25, 2021, claiming priority to March 26, 2020 (see English Translation of description provided), as evidenced by NCBI NP_008982 (printed 2026).
Ueno discloses human patient-derived plasma (a pharmaceutical composition) containing exosomes identified as loaded with Fas-associated factor 1 (FAF1) protein encoded by NCBI accession number NP_008982 (see Example 1; Table 1-1 and Table 4-4, Table 5-4), which is 100% identical to instant SEQ ID NO:1 (see sequence alignment above). Ueno also teaches isolating exosomes in PBS solution with a pH around 7 (a pharmaceutical composition) (see 7th page of English translation, last paragraph).
12. Claim(s) 1, 3, 7-9, 11-13 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by KR102049199, Kim, published November 26, 2019 with English translation of description and claims.
With regard to claims 1, 3, 7-9, Kim teaches isolating exosomes loaded with FAF1 protein in PBS (pharmaceutical composition) (see English translation of description [81-83]; Figure 4), wherein exosomes are sized 30-100 nm (p. 3, [37]).
With regard to claims 11-12, Kim teaches producing FAF1 protein-loaded exosomes comprising introducing a polynucleotide encoding FAF1 protein into cancer (tumor) cells to obtain transformed cells; culturing the transformed cancer cells; and isolating the exosomes from the cells (see English translation of description [81-83]; Figure 4).
With regard to claim 13, Kim teaches a method comprising 1) treating cancer cells having decreased expression of FAF1 protein with a vector encoding FAF1 protein (contacting cancer cells with a test substance), 2) measuring increased levels of FAF1 protein in exosomes from the cells; and 3) identifying the vector encoding FAF1 protein results in increased levels of FAF1 protein in exosomes as compared to cells not treated with the vector (Example 2-4, [80-83]).
The preamble of claim 13: “for screening cancer therapeutic agents” is merely suggested intended use for the claimed method. MPEP 2111.02 states: If the body of a claim fully and intrinsically sets forth all of the limitations of the claimed invention, and the preamble merely states, for example, the purpose or intended use of the invention, rather than any distinct definition of any of the claimed invention’s limitations, then the preamble is not considered a limitation and is of no significance to claim construction.
Claim Rejections - 35 USC § 103
The following is a quotation of 35 U.S.C. 103 which forms the basis for all obviousness rejections set forth in this Office action:
A patent for a claimed invention may not be obtained, notwithstanding that the claimed invention is not identically disclosed as set forth in section 102, if the differences between the claimed invention and the prior art are such that the claimed invention as a whole would have been obvious before the effective filing date of the claimed invention to a person having ordinary skill in the art to which the claimed invention pertains. Patentability shall not be negated by the manner in which the invention was made.
13. Claims 1, 3, 7-9, 15, 18-23 are rejected under 35 U.S.C. 103 as being obvious over US Patent Application Publication 2005/0261190, Lee et al; in view of CN101899443, Jin et al, published 2010 with English translation of description and claims; and KR102049199, Kim, published November 26, 2019 with English translation of description and claims; and Johnsen et al (Biochimica et Biophysica Acta, 2014, 1846:75-87).
Lee teaches a method of treating a subject having cancer that lacks expression of FAF1 ([109]), the method comprising administering a therapeutically effective amount of a pharmaceutical composition comprising FAF1 protein to the subject, because increasing FAF1 protein levels increases apoptosis of cancer cells ([9-11]; [54]; [64]; [84]; [95]; [108-118]; [121-123]);
wherein the cancer includes melanoma, sarcoma, bladder carcinoma, brain tumor, breast cancer, cervical cancer, colorectal cancer, esophageal cancer, endometrial cancer, hepatocellular carcinoma, gastrointestinal stromal tumor (GIST), laryngeal cancer, lung cancer, osteosarcoma, ovarian cancer, pancreatic cancer prostate cancer, renal cell carcinoma or thyroid cancer ([87]; [96]);
wherein the composition is delivered via oral, intravenous, intramuscular, subcutaneous, intranasal, intradermal, intraperitoneal route ([111-117]); and
the method further comprising administering additional therapeutic agents ([41]).
Lee demonstrates that increased levels of human FAF1 protein functions to increase apoptotic death of cancer cells, which suppresses tumor formation, and demonstrates transfecting HEK293 cells to overexpress human FAF1 protein (Examples 4.1 and 4.8). Lee demonstrates that FAF1 is decreased in expression levels in ovarian cancer tissue compared to normal tissue (Example 5; [87]).
Lee suggests administering the protein by known delivery methods including a vesicle, in particular a liposome, to protect the protein from enzymatic digestion and teaches motivation to increase half-life ([64]; [121-123]; [111-112]).
Lee does not teach the FAF1 protein is comprised in, and delivered by, vesicles that are a FAF1 protein-loaded exosomes sized in the range of 50-200 nm, or that target tumor cells (instant claims 1, 3, 7-9, 15, 18-23). Lee does not teach producing FAF1 protein-loaded exosome composition by transfecting cancer cells or HEK-293 cells with FAF1 gene encoding protein, culturing the cells, and isolating the exosomes from the cells (instant claims 11 and 12).
Jin also teaches treating cancer by administering FAF1 protein, demonstrating that increasing FAF1 levels increases apoptosis of cancer cells, including treating cervical, gastric, colon, prostate, and pancreatic cancers (see English translation of description p. 8 fourth aspect). Jin teaches a pharmaceutical composition comprising an effective amount of FAF1 protein (English translation of description at p. 9. sixth and seventh aspects; English translation of claims at claim 9). Jin also teaches that FAF1 apoptosis is inhibited by miR-24, and demonstrates that administering antisense inhibitors of miR-24 successfully increases FAF1 expression in cells and increases apoptosis (Figures 1, 2C, 6 and 7) (see English translation of description p. 2).
Thus, Lee and Jin establish the function of increasing FAF1 protein levels in cancer cells increases apoptosis to treat cancer.
Kim also teaches FAF1 protein is known to induce apoptosis of cells (English Translation of description at [26]; [39-42]) and demonstrates cells secrete FAF1 protein through exocytosis in the form of exosomes, which results in the induction of apoptosis in surrounding cells ([54]). Kim demonstrates transfecting cancer cells with a vector encoding FAF1 protein, expressing the FAF1 protein, and confirming FAF1 protein is secreted through exosomes ([Examples 1-2, [59-83]; Figure 4). Kim demonstrates that cancer cells transfected to express FAF1 protein (donor cells), secrete FAF1 through exosomes, and the donor cells successfully transferred FAF1 protein to recipient cells, resulting in increased FAF1 protein in the recipient cells and inducing apoptosis in both the donor and recipient cells (Example 3; [85-101]; Figures 6 and 7).
Johnsen reviews the established use of exosomes, having a diameter of 30-120 nm (section 1), to deliver protein and other therapeutics to tumors for cancer therapy. Johnsen demonstrates the known process of exosomes secreted from a parent (donor) cell to a recipient cell, where the recipient cell internalizes the exosome to incorporate its cargo (Figure 1). Johnsen reviews several known clinical applications of administering exosomes in cancer therapy to deliver the cargo, particularly protein, loaded within the exosome and the various administration routes (abstract; Figure 2; Tables 1-4; section 4.2 and section 4.5). Johnsen reviews known methods of producing exosomes carrying the therapeutic protein cargo by transfecting donor exosome-producing cells to overexpress a protein product, thereby resulting in exosomes loaded with the protein, and isolating the exosome for administration (section 4.3.3; Table 2; Figure 2). Johnsen reviews several examples of therapeutic exosomes produced in cancer cells by transfection including in HEK-293 cells (Table 2). Johnsen reviews known methods for making exosomes that have an additional function of targeting cancer cells (section 4.4; Table 3). Johnsen teaches an advantage of using exosomes to deliver therapeutics over using liposomes is that exosomes contain exosome membrane proteins that result in better targeting, fusion with, and drug delivery to recipient cells (section 5; Figure 3). Johnsen teaches exosomes are expected to have an advantage of longer half-life in circulation compared to liposomes (section 5).
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed to make and use a pharmaceutical composition comprising exosomes in the size range of 50-200 nm loaded with FAF1 protein for treating cancer in the method of Lee. One would have been motivated to, and have a reasonable expectation of success to, because: (1) Lee and Jin both teach administering FAF1 protein to cancer cells in order to increase FAF1 levels and induce apoptosis, wherein Lee suggests administering the FAF1 protein in a vesicle or liposome for protection and to enhance half-life; (2) Lee, Jin, and Kim all recognize that FAF1 functions to induce apoptosis, where Kim demonstrates FAF1-transfected cells successfully produce exosomes loaded with FAF1 protein and the exosomes successfully increase FAF1 protein and induce apoptosis in recipient cells; and (3) Johnsen teaches exosomes are sized 30-120 nm; are an established and successful vesicle of delivery for protein therapeutics; and teaches the advantages of exosomes over liposome delivery including better targeting, fusion with, and drug delivery to recipient cells, and longer half-life.
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed to produce exosomes that target tumor cells for cancer treatment. One would have been motivated to, and have a reasonable expectation of success to, because: (1) Lee and Jin both teach administering FAF1 protein to cancer cells in order to increase FAF1 levels and induce apoptosis in cancer cells; and (2) Kim teaches and demonstrates exosomes can be produced to target tumor cells for efficient delivery of their therapeutic cargo.
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed to make the exosomes loaded with FAF1 by transfecting cancer or HEK293 cells with a polynucleotide encoding FAF1 protein; culturing the cells, and isolating the exosomes from the cells. One would have been motivated to, and have a reasonable expectation of success to, because: (1) Johnsen teaches methods of producing exosomes by transfecting cells to overexpress the therapeutic protein cargo and isolating the exosomes for administration is an established method, and teaches this method is already successfully practiced using exosome-producing HEK-293 cells; (2) Lee demonstrates successfully transecting HEK-293 cells to overexpress human FAF1 protein; and (3) Kim demonstrates successfully producing FAF1-loaded exosomes by transfecting cancer cells to overexpress FAF1.
14. Claim(s) 2, 10, and 17 are rejected under 35 U.S.C. 103 as being unpatentable over US Patent Application Publication 2005/0261190, Lee et al; CN101899443, Jin et al, published 2010 with English translation of description and claims; and KR102049199, Kim, published November 26, 2019 with English translation of description and claims; and Johnsen et al (Biochimica et Biophysica Acta, 2014, 1846:75-87); as applied to claims 1, 3, 7-9, 15, 18-23 above, and further in view of WO 2021/193802, Ueno et al, filed March 25, 2021, claiming priority to March 26, 2020 (see English Translation provided), as evidenced by NCBI NP_008982 (printed 2026).
Lee, Jin, and Kim (the combined references) teach making and using a pharmaceutical composition comprising FAF1 protein-loaded exosomes for the treatment of cancer, as set forth above.
The combined references do not teach the FAF1 protein comprises instant SEQ ID NO:1.
Ueno discloses human cancer patient-derived plasma (a pharmaceutical composition) containing exosomes identified as loaded with Fas-associated factor 1 (FAF1) protein encoded by NCBI accession number NP_008982 (see Example 1; Table 1-1 and Table 4-4, Table 5-4), which is 100% identical to instant SEQ ID NO:1 (see sequence alignment above). Ueno also teaches isolating exosomes in PBS solution with a pH around 7 (a pharmaceutical composition) (see 7th page of English translation, last paragraph).
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed for the FAF1 protein of the combined references to comprise SEQ ID NO:1 (NP_008982). One would have been motivated to, and have a reasonable expectation of success to, because the combined references teach utilizing human FAF1 protein associated with cancer as the therapeutic protein contained in the exosomes, and Ueno teaches the sequence of human FAF1 protein associated with cancer is known as SEQ ID NO:1 (NP_008982).
15. Claim 13 is rejected under 35 U.S.C. 103 as being obvious over US Patent Application Publication 2005/0261190, Lee et al; in view of CN101899443, Jin et al, published 2010 with English translation of description and claims; and KR102049199, Kim, published November 26, 2019 with English translation of description and claims.
Lee teaches as set forth above. Lee further suggests screening for a substance that induces apoptosis and cell death ([2]) and demonstrates transfecting cancer cells with a vector encoding human FAF1 protein, detecting increased FAF1 protein levels compared to control-treated cells, and identifying the vector encoding FAF1 protein as capable of increasing FAF1 protein levels in the cell and inducing apoptosis and cancer cell death (Examples 2-4).
Lee does not teach conducting a screening assay for test substances that function as a cancer therapeutic agent, the assay comprising contacting a test substance with cancer cells having decreased expression of FAF1, measuring the level of FAF1 protein in exosomes from the cells, and selecting the test substance that increases levels of FAF1 protein in the exosomes as compared to untreated cells.
Jin teaches as set forth above. Jin further teaches screening for test substances that treat cancer having reduced expression of FAF1, and exemplifies contacting HEK293, HeLa, or gastric cancer cells with miR-24 antisense/inhibiting test substance, detecting increased FAF1 protein expression in the cells, and inducing apoptosis and cell death in the cancer cells, unlike in untreated controls (English translation of description at p. 3; Figures 1-7 p. 3-4; English translation of claims at claim 10). Jin teaches suitable cells for the invention include HeLa cells (paragraph spanning p. 6-7). Jin teaches selecting miR-24 inhibitors, such as antisense, to prepare a treatment for cancer with abnormal expression of FAF1 gene (p. 8). Jin teaches an in vivo model of drug screening comprising contacting a animal model having a tumor with reduced FAF1 expression, administering a drug to be screened to the animal, and determining whether the drug causes tumor regression or cure the animal (p. 9).
Kim teaches as set forth above. Kim further teaches a screening method to identify test substances that reduce or do not reduce FAF1 protein in exosomes, the method comprising contacting cells in vitro with a test substance, measuring FAF1 in exosomes form the cells, and determining if the test substance inhibits FAF1 protein levels as compared to cells not treated with the test substance (see English translation [12-15]; [23-25]; [33-36]; [54]). Kim exemplifies transfecting cancer cells under-expressing FAF1 with a vector encoding FAF1 protein (contacting cancer cells with a test substance), measuring increased levels of FAF1 protein in exosomes from the cells as compared to cells not treated with the vector (Example 2-4, [80-83]). Kim exemplifies transfecting cancer cells under-expressing FAF1 with a vector encoding FAF1 protein, contacting the cells with either GW4869 or Monensin test substances, determining Monensin increased FAF1 protein secretion as compared to the untreated control group, and determining GW4869 decreased FAF1 protein secretion as compared to an untreated control group ([Example 2-3; [75-78]).
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed to screen for cancer therapeutic agents comprising contacting a test substance with cancer cells having decreased expression of FAF1, measuring the level of FAF1 protein in exosomes isolated from the cells, and selecting the test substance in which the level of FAF1 protein is increased as compared to an untreated control group in the method of Lee. One would have been motivated to, and have a reasonable expectation of success to because: (1) Lee and Jin establish that anticancer agents function to kill cancer cells by increasing levels of FAF1 protein and inducing apoptosis; (2) Lee and Jin suggest screening for anti-cancer agents that increase FAF1 protein in cancer cells and induce apoptosis; (3) Jin demonstrates successfully identifying an anticancer agent that increases FAF1 protein level in cancer cells and induces apoptosis by contact the cancer cells with a test agent and selecting the agent as a cancer therapeutic that increases FAF1 protein in the cancer cells as compared to untreated controls; and (4) Kim teaches and demonstrates that increased FAF1 in cancer cells is also increased in their secreted exosomes, and successfully teaches/demonstrates screening assays comprising contacting cancer cells with a test agent to determine if the agent increases or decreases FAF1 protein in exosomes isolated in the cells compared to untreated controls.
16. Claim(s) 14 is rejected under 35 U.S.C. 103 as being unpatentable over US Patent Application Publication 2005/0261190, Lee et al; CN101899443, Jin et al, published 2010 with English translation of description and claims; and KR102049199, Kim, published November 26, 2019 with English translation of description and claims; and Johnsen et al (Biochimica et Biophysica Acta, 2014, 1846:75-87); as applied to claim 13 above, and further in view of Johnsen et al (Biochimica et Biophysica Acta, 2014, 1846:75-87).
Lee, Jin, and Kim (the combined references) teach a method for screening for cancer therapeutic agents comprising: (1) treating cancer cells having decreased FAF1 expression with a test substance; (2) measuring the level of FAF1 protein in exosomes isolated from the cells in step (2); and (3) selecting the test substance in which the level of FAF1 protein in step (2) is increased as compared with a control group is not treated with the test substance, as set forth above.
Jin further demonstrates contacting HeLa cells with a test substance (miR-24 antisense and staurosporin) and determining that test agents induced apoptosis in the HeLa cells (Figure 5; English Translation of description at page 4), wherein Jin established that mi-R24 antisense increases FAF1 protein levels in cells to induce apoptosis, as stated above.
The combined references do not teach utilizing HeLa cells in the screening assay.
Johnsen teaches as set forth above. Johnsen further teaches that numerous cell types can be used as exosome factories and the most commonly used include HEK-293 and HeLa cells (section 4.1).
It would have been prima facie obvious to one of ordinary skill in the art at the time the invention was filed to utilize HeLa cells in the screening assay of the combined references to identify anticancer test substances that increase FAF1 protein in secreted exosomes as compared to untreated cells. One would have been motivated to, and have a reasonable expectation of success to, because: (1) the combined references demonstrate successfully using a variety of different cancer cells in screening assays to identify agents that increase FAF1 protein and induce apoptosis; (2) Jin teaches HeLa cells are successfully killed by contacting them with an agent that is known to increase FAF1 protein level in cells; and (3) Johnsen teaches HeLa cells are commonly and successfully used to produce exosomes.
17. Conclusion: No claim is allowed.
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/Laura B Goddard/ Primary Examiner, Art Unit 1642