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 .
Restriction/Election
Applicant’s election of Group I, drawn to a method for extending the shelf life of donor blood containing red blood cells (RBC), and claims 1-12 encompass the elected invention, in the reply filed on 08/06/2025 is acknowledged. Because applicant did not distinctly and specifically point out the supposed errors in the restriction requirement, the election has been treated as an election without traverse (MPEP § 818.01(a)).
Claims 1-27 are pending.
Claims 13-27 are withdrawn from further consideration pursuant to 37 CFR 1.142(b) as being drawn to nonelected inventions, there being no allowable generic or linking claim.
Claims 1-12 are currently under examination.
Priority
This application 17919364 filed on 10/17/2022 is a national phase application under 35 U.S.C. § 371 that claims priority to International Application No. PCT/US2021/029104 field on 04/26/2021, which claims priority to U.S. Provisional Patent Application Serial No. 63/016,395, field on 04/28/2020.
A certified copy of priority document International Application No. PCT/US2021/029104 field on 04/26/2021 has been submitted of the record by Applicants on 10/17/2022.
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.
Claim 12 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 12 recites "the mediation" in line 1 of the limitation “wherein the mediation is determined by measuring the membrane integrity of the cyclodextrin composition-treated REC as compared to non-cyclodextrin composition-treated RBC". There is insufficient antecedent basis for this limitation in the claim 1.
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.
This application currently names joint inventors. In considering patentability of the claims the examiner presumes that the subject matter of the various claims was commonly owned as of the effective filing date of the claimed invention(s) absent any evidence to the contrary. Applicant is advised of the obligation under 37 CFR 1.56 to point out the inventor and effective filing dates of each claim that was not commonly owned as of the effective filing date of the later invention in order for the examiner to consider the applicability of 35 U.S.C. 102(b)(2)(C) for any potential 35 U.S.C. 102(a)(2) prior art against the later invention.
Claims 1-7, 9, 11 and 12 are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Motoyama et al. (2006) (Motoyama et al., Effect of 2,6-di-O-methyl-acyclodextrin on hemolysis and morphological change in rabbit's red blood cells, Eur J Pharm Sci., 2006 Oct 1;29(2):111-9, doi: 10.1016/j.ejps.2006.06.002. Epub 2006 Jun 10; This reference is cited in the Restriction/Election mailed on 06/17/2025).
Claim interpretations: The limitations “extending the shelf life of donor blood containing red bold cells (RBC)” recited in the preamble of claim 1, “iii) extending the shelf life of the donor blood and/or RBC of step ii),” recited in claim 1, and “iv) preserving the cyclodextrin-treated donor blood and/or RBC of step iii) in storage” recited in claim 2 are considered as the consequences of performing steps i) and ii) recited in claim 1.
Regarding claims 1-4, 9, and 12, Motoyama et al. (2006) teaches that "The effects of 2,6-di-O-methyl-a-cyclodextrin (DM-a-CyD) [which reads on the limitation recited in instant claim 3] on hemoolysis and morphological changes in rabbit's red blood cells (RBC) [which reads on the limitation “mammalian blood” recited in instant claim 9] were examined, compared with those of a-cyclodextrin (CyD) and 2-hydoxypropyl-a-cyclodextrin (HP-a-CyD). The hemolytic activity of a-CyDs increased in the order of HP-a-CyD < a-CyD < DM-a-CyD. The three a-CyDs induced morphological changes of RBC from discocyte to stomatocyte. At the same concentration (3 mM) of a-CyDs, DM-a-CyD and a-CyD released phospholipids, rather than cholesterol, and DM-a-CyD markedly released proteins from RBC membranes, compared to a-CyD and HP-a-CyD. The treatment of RBC with DM-a-CyD lowered the extent of a fluorescent sphingomyelin analogue from lipid rafts of RBC membranes in a concentration-dependent manner [which reads on the limitation “measuring the membrane integrity” recited in instant claim 12]. These results suggest that DM-a-CyD has higher hemolytic and morphological change activity than a-CyD and HP-a-CyD through more extraction of phospholipids including sphingomyelin [which reads on the limitation “sphingomyelin (SM)” recited in instant claim 4] and proteins, not cholesterol, from RBC membranes than a-CyD and HP-a-CyD. (See ABSTRACT). Motoyama et al. (2006) further teaches that "In brief, 2 ml of rabbit blood was incubated with 12.5 mg/ml of NBD-sphingomyelin [which reads on the limitation “exogenous lipid” recited in step ii) instant claim 1] at 4°C for 30 min. The blood was centrifuged at 1000 x g for 5min and the pellet was washed three times with cold PBS. Two milliliters of the a-CyDs solutions were added to 1ml of the RBC suspension." (See section 2.5, right column, page 112).
Regarding claims 5-7, Motoyama et al. (2006) teaches in section 2.2. Hemolytic activity that “RBC were isolated from Japanese white male rabbits (Kyudo, Tosu, Japan) as described previously (Uekama et al., 1981; Irie et al., 1982). Isolated RBC were centrifuged at 1000×g for 5min and washed three times with 10mM of phosphate buffered saline (PBS, pH 7.4). Five percent of RBC suspension in PBS was incubated with 2 ml of PBS (pH 7.4) containing a-CyDs for 30 min at 37°C. After centrifugation (1000×g for 10 min) the optical density of the supernatant was measured at 543 nm. Results were expressed as percent of total hemolysis, which was obtained when RBC were incubated in water only. All hemolytic assays were carried out on the same day of blood collection (See left column, page 112).
Regarding claim 11, Motoyama et al. (2006) teaches RBC treated with 3 mM and 5 mM DM-a-CyD showed stomatocytes and spherostomatocytes, respectively (Fig. 2D). Similar results were observed in RBC treated with a-CyD at the concentrations of 6 mM and 12.5 mM (Fig. 2B) (See right column, page 113).
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Fig. 2 – (A–D) Scanning electron microscopic analysis of morphological changes in RBC induced by a-CyDs. RBCs were treated with a-CyDs using the same methods as used for hemolytic assay. After fixation of samples with 2% (v/v) glutaraldehyde, RBC were resuspended with distilled water. The samples were dried, and then were sputtered with gold using an Ion Coater IB-3, followed by observation using a Hitachi S-510 SEM. These pictures show representative data for three experiments.
Motoyama et al. (2006) teaches that “In conclusion, we revealed that DM-a-CyD has hemolytic activity stronger than a-CyD and HP-a-CyD. DM-a-CyD was found to induce the morphological change to stomatocyte through the selective extraction of phospholipids including
sphingomyelin and proteins, rather than cholesterol, from RBC membranes. These results will provide useful information for the pharmaceutical and cell biological applications of methylated CyD derivatives.” (See left column, page 118).
Claim Rejections - 35 USC § 103
In the event the determination of the status of the application as subject to AIA 35 U.S.C. 102 and 103 (or as subject to pre-AIA 35 U.S.C. 102 and 103) is incorrect, any correction of the statutory basis (i.e., changing from AIA to pre-AIA ) for the rejection will not be considered a new ground of rejection if the prior art relied upon, and the rationale supporting the rejection, would be the same under either status.
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.
The factual inquiries for establishing a background for determining obviousness under 35 U.S.C. 103 are summarized as follows:
1. Determining the scope and contents of the prior art.
2. Ascertaining the differences between the prior art and the claims at issue.
3. Resolving the level of ordinary skill in the pertinent art.
4. Considering objective evidence present in the application indicating obviousness or nonobviousness.
Claims 1-12 are rejected under 35 U.S.C. 103 as being unpatentable over in view of Motoyama et al. (2006) (Motoyama et al., Effect of 2,6-di-O-methyl-acyclodextrin on hemolysis and morphological change in rabbit's red blood cells, Eur J Pharm Sci., 2006 Oct 1;29(2):111-9, doi: 10.1016/j.ejps.2006.06.002. Epub 2006 Jun 10) in view of Stoll et al. (2011) (Stoll et al., Liposomes alter thermal phase behavior and composition of red blood cell membranes, Biochim Biophys Acta, 2011 Jan;1808(1):474-81. doi: 10.1016/j.bbamem. 2010.09.012. Epub 2010 Sep 29).
The teaching of Motoyama et al. (2006) have been documented above in the rejection of claims 1-7, 9, 11 and 12 under 35 U.S.C. 102(a)(1) as being anticipated by Motoyama et al. (2006).
Motoyama et al. (2006) does not explicitly teach (i) the limitation regarding extending the shelf life of donor blood containing red blood cells (RBC) recited in instant claim 1 via cyclodextrin treatment induced alteration of the ratio of cholesterol/phospholipids in lipid bilayers of RBC membrane, (ii) the limitation regarding extended RBC shelf life recited in instant claim 8, and (iii) the limitation regarding the red blood cells (RBCs) are from human blood recited in instant claim 10.
(i)-(ii) Regarding extending the shelf life of donor blood containing red blood cells (RBC), Stoll et al. (2011) teaches that “There is an urgent need for the development of red blood cell (RBC) bio-preservation techniques that maintain in vitro RBC function to ensure permanent availability of all RBC types. The earliest and most widely investigated approach to RBC bio-preservation is hypothermic storage, which allows storage of blood for up to 42 days at 1–6 °C in preservative solutions [which is relevant to the limitation recited in instant claim 8, but in the absence of cyclodextrin treatment]. Cryopreservation of RBCs can be done using glycerol as cryoprotective agent. Membranes are one of the primary sites of injury during hypothermic storage, cryopreservation or freeze-drying of mammalian cells. The stabilizing effect of liposomes on sperm during bio-preservation is known for some time. Recently, liposomes have been used as protective agents for cryopreservation and freeze-drying of red blood cells. These studies have established the beneficial effect of liposomes for bio-preservation of cells. How precisely liposomes interact with cells and stabilize bio-membranes, remains hitherto unknown. The protective properties of liposomes can likely be attributed to stabilizing membrane modifications due to lipid and cholesterol transfer between liposomes and cells. Cholesterol can exchange rapidly between different membrane bilayers, whereas lipid transfer is a relatively slow process.” (See Introduction, Bridging paragraph from left to right column, page 474).
Relevant to the limitations recited in claims 5-7, “wherein the donor blood and/or RBCs are exposed to the cyclodextrin composition for a period of time sufficient to reconstitute the lipids in the outer leaflet of the membranes of the RBC in the donor blood”, in addition to the teachings “Five percent of RBC suspension in PBS was incubated with 2 ml of PBS (pH 7.4) containing a-CyDs for 30 min at 37°C” by Motoyama et al. (2006), Stoll et al. (2011) discloses related teachings in section “2.4. Cholesterol extraction by methyl-ß-cyclodextrin: The RBC concentrate was diluted 1:10 in HBSI buffer containing 10 mM methyl-β-cyclodextrin (Sigma Aldrich, St. Louis, MO) and incubated for 70 min at 37 °C. The cholesterol depleted RBCs were washed in HBSI buffer”. Stoll et al. (2011) further teaches the alteration of the ratio of cholesterol/phospholipids in lipid bilayers of RBC membrane induced by cyclodextrin; and in this regard, Stoll et al. (2011) teaches in section 3.3 that “Cholesterol depletion with methyl-β-cyclodextrin: In order to study the effect of cholesterol on RBC membrane phase behavior, methyl-β-cyclodextrin (MβCD) was used to extract cholesterol from the cellular membranes. Methyl-β-cyclodextrin is an efficient agent for removing cholesterol from cell membranes in a short procedure. Fig. 5 shows a wave number vs. temperature plot of RBC ghosts prepared from RBCs that were treated with 10 mM MβCD. The MβCD-treated cells display an increase in membrane conformational disorder at supra-zero temperatures similarly to DLPC treated RBCs. The DLPC-treated RBCs, however, display a greater increase in membrane conformational disorder in the supra-zero temperature regime compared to the MβCD-treated cells”. (See right column, page 477).
(iii) Regarding claim 10, Stoll et al. (2011) teaches in section 2.2. Pretreatment of blood: “Human blood was obtained from the Institute for Transfusion medicine (Hannover Medical School). Venous blood was collected from healthy adults, with informed consent, according to institutional protocols. Blood was anticoagulated with citrate phosphate dextrose adenine (CPDA). Whole blood was centrifuged at 4000 ×g for 10min. The buffy coat and supernatant were discarded, and the RBC fraction was employed for experiments. Anticoagulated blood was stored in polypropylene Falcon tubes at 1–6 °C until used for experiments”. (See left column, page 475).
It would have been prima facia obvious for a skilled artisan to incorporate the teachings of Stoll et al. (2011) into the teachings of Motoyama et al. (2006) to reach the method recited in instant claim 1 regarding extending the shelf life of red blood cells with reasonable expectation of success because both Stoll et al. (2011) and Motoyama et al. (2006) are directed to the effect of cyclodextrin on the structural alteration of mammalian red blood cell membranes. Moreover, (i) Motoyama et al. (2006) concludes that “These results will provide useful information for the pharmaceutical and cell biological applications of methylated CyD derivatives.” (See left column, page 118); and (ii) Stoll et al. (2011) discloses that “The protective properties of liposomes can likely be attributed to stabilizing membrane modifications due to lipid and cholesterol transfer between liposomes and cells” (See Introduction, page 474 of Stoll et al.)
A skilled artisan would have been motivated to incorporate the teachings of Stoll et al. (2011) into the teachings of Motoyama et al. (2006) because (i) Stoll et al. (2011) specifically teaches that “DPPC/cholesterol liposomes and DMPC liposomes were found to be protective during freezing and freeze-drying, respectively. Protective properties of liposomes can sometimes be enhanced by adding cholesterol to the liposomes. DLPC/cholesterol liposomes, for example, have been shown to increase sperm motility after freezing and thawing, whereas pure DLPC liposomes decreased motility after thawing” (See right column page 480), and (ii) Motoyama et al. (2006) discloses that “DM-a-CyD may induce morphological changes of RBC
through altering the ratio of cholesterol/phospholipids in lipid bilayers of RBC membrane (See left column, page 117, and Fig 4 on page 116).
Conclusion
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/WU CHENG W SHEN/Supervisory Patent Examiner, Art Unit 1682