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 .
Information Disclosure Statement
The submitted information disclosure statement (IDS) was filed on 02/24/2023. The submission is in compliance with the provisions of 37 CFR 1.97. Accordingly, the information disclosure statement is being considered by the examiner.
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 11 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 11 is indefinite because it recites: “11. The method according to claim 1, CHARACTERIZED in that it further comprises adjusting the pH of the step mixture to between 9 and 10.”, wherein which step is not defined. For examining purposes, examiner will assume that dissolving bovine serum albumin in ultra-pure water to prepare solution B, adding surfactant, using NaOH solution to adjust the pH value is 8-10; adding the solution A into the solution B, mixing the solution C. See the rejection of Claim 11 by Zhang below. Appropriate correction is required.
Claim Rejections - 35 USC § 103
4. 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 non-obviousness.
Claim(s) 1-10, 12-13, 14, 15-16, 18-21 and 22 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Nosrati et al. (Nosrati et al., Bovine Serum Albumin (BSA) coated iron oxide magnetic nanoparticles as biocompatible carriers for curcumin-anticancer drug. Bioorganic Chemistry 76 (2018) 501–509) in view of KR 101125232 B1.
Claim(s) 1-10, 12-13 and 19-21,
Nosrati et al. teach a simple improved method for the preparation of magnetic albumin based nano-carrier. (pg. 502, left col., 2nd par.). In this method, a desolvation agent such as acetone or ethanol can add to the aqueous solution of albumin under a constant stirring condition using a magnet. (pg. 502, left col., 1st par.).
To an aqueous solution (32 mL in deionized water) of a mixture of FeCl3.6H2O (0.55 g) and FeCl2.4H2O (0.2 g), BSA (2 g) was added and kept at room temperature for 15 min under vigorous stirring.
Then, 64 mL acetone was added to above solution drop wise at a rate of 2 mL/min under constant stirring at room temperature.
For the stabilization of the unstable particles, 40 mg EDC was added for cross linking. The stirring condition was continued for 2 h to ensure the cross linking of all amino acid residues. Then the temperature was increased to 50°C. Next, under vigorous stirring and N2 gas a solution of ammonium hydroxide (15 mL NH4OH (25%)) was added till the pH was raised to ~11 at which a black suspension was formed.
This suspension was then stirred at room temperature for 3 h. The resulting BSA coated magnetic nanoparticles (F@BSA NPs) were purified by three cycles of centrifugation at 18,000 rpm for 15 min and washed with deionized water several times and dried in a vacuum oven overnight. (pg. 502, right col., 2nd par.).
A mixture of FeCl3.6H2O (0.55 g) and FeCl2.4H2O (0.2 g), BSA (2 g), or the ratio of BSA 2 g over iron salt forms 0.75 g is 10:3.75.
64 mL acetone was added to above solution of 2g albumin, or 64 mL/2000 mg albumin,
Nosrati et al. teach a, b, c and d steps, but do not have the stirring speed of a rotor/stator-type homogenizer.
KR 101125232 B1 teach a biocompatible polymer microsphere equipped with super paramagnetic iron oxide nanoparticles (SPIOs). Superparamagnetic Iron Oxide Nanoparticles, Microspheres, and Polymers. (Abs). One of the biocompatible polymers solutions is albumin, to be mixed with a solvent, which may be used, and a person skilled in the art may select and use a suitable solvent according to the type of biocompatible polymer. (pg. 3, 3rd last par.).
The microspheres prepared according to the production method of the present invention are spherical with a ratio of long axis to short axis of 1: 0.9 to 1.1, specifically 1: 0.95 to 1.05.nanoparticles;
(a) uniformly mixing the biocompatible polymer solution with the superparamagnetic iron oxide nanoparticles;
(b) dropping the biocompatible polymer solution and the superparamagnetic iron oxide nanoparticle mixture into a mixture consisting of an organic solvent upper layer and a coacervation medium lower layer which are not mixed with water to form microspheres;
(c) cross-linking the formed microspheres;
(d) washing the crosslinking agent left unreacted with a solvent such as acetone and distilled water and drying under conditions such as lyophilization or dry oven, room temperature, etc. to provide. (pg. 5, 1st par.).
The superparamagnetic iron oxide nanoparticles and the biocompatible polymer solution may be mixed to uniformly mix the iron oxide nanoparticles at several tens of thousands of rpm using a stirrer, a mixer, a homogenizer, and the like. (pg. 4, 1st par.).
In the crosslinking reaction, the degree of crosslinking may be controlled by adjusting the concentration and reaction time of the crosslinking solution, and the degree of crosslinking according to the crosslinking time may be varied according to the type of the crosslinking agent. (pg. 5, 2nd last par.).
Nosrati et al. teach step a, b and c: vigorous mixing, constant stirring and vigorous stirring.
KR 101125232 B1 teach mixed to uniformly mix the iron oxide nanoparticles at several tens of thousands of rpm using a stirrer, a mixer, a homogenizer, and the like. (pg. 4, 1st par.).
A stirring speed of 8,000 to 20,000 RPM means vigorous stirring. This speed range is exceptionally high for standard lab magnetic stirrers and is typically associated with specialized equipment like overhead stirrers or high-shear mixers. The effect of stirring speed depends on
Viscosity: A highly viscous (thick) liquid will require a much higher RPM to achieve the same mixing effect as a low-viscosity liquid like water.
Impeller type and size: The specific geometry and size of the stirring impeller or stir bar have a significant effect on the mixing efficiency.
Container size and geometry: The size and shape of the mixing vessel impact the fluid dynamics and the resulting turbulence.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the invention to prepare albumin nanoparticles loaded with magnetic nanoparticles by the steps taught by Nosrati et al. with a with vigorous mixing, constant stirring and vigorous stirring in all steps a, b, and c, with the weight ratio 10:3.75 of albumin over iron salts taught by Nosrati et al., and superparamagmetic iron-oxide nanoparticles (SPIONs), mixing at several tens of thousands of rpm by homogenizer taught by KR 101125232 B1, since together they have proven they could successfully prepare these magnetic nanoparticles.
With regard to claim 14,
Nosrati et al. teach 64 mL acetone was added to above solution, of FeCl3.6H2O (0.55 g) and FeCl2.4H2O (0.2 g), BSA (2 g) in 32 mL, of drop wise at a rate of 2 mL/min under constant stirring at room temperature, which is 64 mL acetone into 2 g BSA or 0.032 mL acetone into 1mg BSA. This is 10-fold lower than the applicant limitation in the claim.
It would have been obvious to one of ordinary skill in the art before effective filing date of the invention to vary ratio of acetone over BSA, with different mixer, like magnetic stirrer or homogenizer, or rotor/stator homogenizer, to have and how much water in the mixture in the step a and to obtain F@BSA@CUR NPs on the smaller average size of 56 ± 11.43 nm. (Abs) taught by Nosrati et al. or to have less concentrated BSA 10 mg/mL in 50 mL of distilled water and less iron concentration in the BSA particles at 1, 2, or 3 mg of superparamagnetic iron-oxide nanoparticles (SPION), and then higher acetone 0.3-0.7 mL acetone/mg of BSA into 500 mg BSA at concentration 10mg/mL. (SPEC, Example 3, pg. 15). There are different parameters to be varied here depending on the one with skill in the art’s goals they want to achieve, higher or lower superparamagnetic iron-oxide nanoparticles (SPION) in BSA to have lower or higher ratios of desolvation agent in albumin, which can be optimized with their goals.
With regard to claim 15 and 16,
Glutaraldehyde is a homobifunctional reagent because it is an organic compound with two identical aldehyde groups that can react with nucleophiles, such as the amino groups on proteins, to form stable cross-links. This dual reactivity makes it an effective tool for stabilizing and linking molecules in various biochemical applications, including protein cross-linking, enzyme immobilization, and histology/cytochemistry.
Nosrati et al. teach after finishing the adding of desolvation agent, a cross-linkers such as
glutaraldehyde or EDC must be added to stabilize the nanoparticles by crosslinking of amino acids residues in the protein. (pg. 502, left col., 1st par.).
KR 101125232 B1 teach in step (c), the crosslinking of the microspheres mixed with the superparamagnetic iron oxide nanoparticles is a synthetic crosslinking reagent glutaraldehyde. (pg. 5, 4th last par.).
With regard to claim 18,
Nosrati et al. teach F@BSA@CUR NPs were spherical in shape with an average size of 56 ± 11.43 nm (mean SD (n = 33)). (Abs).
With regard to claim 22,
Nosrati et al. teach d. the resulting BSA coated magnetic nanoparticles (F@BSA NPs) were purified by three cycles of centrifugation at 18,000 rpm for 15 min and washed with deionized water several times and dried in a vacuum oven overnight. (pg. 502, right col., 2nd par.).
The relationship between round per minute (RPM) with the radius of the rotor r in centimeters and relative centrifugal force (RCF) in g is described by the following formula:
RCF= RPM2 x 1.118 10-5 x r
RCF = 11.2 x Radius x (RPM/1,000)2
https://www.sigmaaldrich.com/US/en/support/calculators-and-apps/g-force-calculator
When the Radius of rotor (cm) is 3, 5 or 11cm; Relative Centrifugal Force (RCF) or g force is 10,867, 18, 112 or 39,846
Claim(s) 1 and 11 and 17 is/are rejected under 35 U.S.C. 102(a)(1) as being anticipated by Nosrati et al. (Nosrati et al., Bovine Serum Albumin (BSA) coated iron oxide magnetic nanoparticles as biocompatible carriers for curcumin-anticancer drug. Bioorganic Chemistry 76 (2018) 501–509) in view of KR 101125232 B1, as described in claim 1 above and further in view of Zhang (CN 103611172 B) and Zhang et al. (CN 104546741 A).
The teachings of Nosrati et al. and KR 101125232 B1 are described in claim 1 above.
Claim 11,
KR 101125232 B1 does not teach the pH of the step mixture. Nosrati et al. teach the pH of the step mixture around 11, which is needed to be clearer that it could be in the range of 9-10.
Zhang et al. (CN 103611172 B) teach a loading-yellow magnetic albumin nano-sphere and preparation method thereof, comprising the following steps: using the improved chemical coprecipitation method for preparing ferro/ferric oxide nano-particle and nano-yellow, using the removing solution-crosslinking method to prepare loading nanometer yellow magnetic albumin nano-sphere. the method is simple and easy, albumin nano-sphere heat therapy, chemotherapy, thermal release, magnetic targeting integrally, so it develops a new path for the treatment of tumor. the new-type nano-carrier as a therapeutic carrier, also can be used for in vivo magnetic resonance imaging research and clinical application research provides reliable experimental research data and theoretical basis. (Abs). Technical solution: loading nanometer yellow magnetic albumin nano-sphere of the invention preparation method, comprising the following steps: (0007). According to mass ratio of 2.5:1 to 6:1, weighing the bovine serum albumin, Fe3O4 magnetic nanoparticle, mix them into de-ionized water, configured to bovine serum albumin concentration is from 10 mg/ml to 24 mg/ml of mixed liquid, according to nanometer yellow with Fe3O4 magnetic nanometer particle quality ratio is 1:200, the nanometer yellow solution into the mixed solution, fully stirring and mixing, adjusting pH value to be alkaline, under the magnetic force stirring slowly adding absolute ethyl alcohol, until the turbid solution, then slowly adding glutaric dialdehyde curing, stirring, pouring the reaction liquid into the centrifugal tube in ultra-high-speed centrifugation to obtain loading-yellow magnetic albumin nano-sphere after washing by using deionized water. (0008).
Zhang et al. (CN 104546741 A) teach 2-methoxy estradiol albumin nano-preparation method, which effectively solves the 2 estradiol albumin nano lyophilized preparation, and realize the preparation application problem of the medicine for treating tumor. method is that the 2-methoxy estradiol is dissolved in the organic solvent anhydrous ethanol to prepare solution A; dissolving bovine serum albumin in ultra pure water to prepare solution B, adding surfactant, using NaOH solution to adjust the pH value is 8-10; adding the solution A into the solution B, mixing the solution C, adding glutaraldehyde to the mixed solution C, stirring uniformly, standing and curing, steaming and removing organic solvent, adding freeze-dry protecting agent in the nanometer solution D into nano solution D, freezing and drying to obtain the product, the method of the invention is simple, easy to produce, good stability, long drug effect, strong targeting, can protect medicine from environment influence, fast effect, small side effect, few administration times, high bioavailability, good stability and simple preparation. (Abs). Zhang et al. (CN 104546741 A) do not teach preparation of magnetic albumin nanoparticles, but teach preparation of albumin nanoparticles, with the pH range 8-10, which can be applied for preparation of magnetic albumin nanoparticles.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the invention to prepare albumin nanoparticles loaded with magnetic nanoparticles by the steps taught by Nosrati et al. with a with vigorous mixing, constant stirring and vigorous stirring in all steps a, b, and c, with the weight ratio 10:3.75 of albumin over iron salts taught by Nosrati et al., and superparamagmetic iron-oxide nanoparticles (SPIONs), mixing at several tens of thousands of rpm by homogenizer taught by KR 101125232 B1, and to have step mixture pH adjusting to be alkaline, which is in the range of 8-12, taught by Zhang et al. (CN 103611172 B) and pH 8-10 taught by Zhang et al. (CN 104546741 A), since together they have proven they could successfully prepare these magnetic nanoparticles.
Claim 17,
Nosrati et al. teach a cross-linkers such as glutaraldehyde or EDC, but do not teach the glutaraldehyde has a concentration at 6.25% and is added at a rate of 10 µl per mg albumin.
KR 101125232 B1 teach a cross-linkers such as glutaraldehyde or others, but do not teach the glutaraldehyde has a concentration at 6.25% and is added at a rate of 10 µl per mg albumin.
Zhang et al. (CN 103611172 B) teach precisely weighing proper amount of bovine serum albumin (250 mg), magnetic powder (Fe3O4 magnetic nano particle) 100 mg, dropping 500Ug-yellow and the
mixing liquid, adding distilled water until the total volume is 25 ml, at this concentration of bovine serum albumin is 10 mg/ml, stirring and mixing uniformly, adjusting the pH
value to alkalescence, slowly adding absolute ethyl alcohol under magnetic
stirring until the solution became turbid and then slowly adding glutaric dialdehyde curing according to Fe3O4 magnetic nanometer particle and 2.5% of glutaraldehyde water solution ratio of 2000 mg/ml, stirring (0026)
Zhang et al. (CN 104546741 A) teach the invention 2- estradiol albumin nano lyophilized preparation method comprising the following steps: (I) ), the 2- estradiol (2-ME) is dissolved in the organic solvent of anhydrous ethanol, 2--methoxy estradiol with weight concentration of l-5mg/ml solution A, (2), bovine serum albumin (BSA) was dissolved in ultrapure water to prepare solution B of weight concentration of the bovine serum albumin 20-60mg/ml. of solution B, stirring and mixing uniformly, regulating the pH value to 8-10 with NaOH solution, (3); the solution A is 0. 05-1. flow rate of /Min; into the solution B, the volume ratio of the solution A and the solution B is 2-3: 1, forming mixed solution C, (4) C adding crosslinking agent in the mixed solution with mass concentration of 8% glutaraldehyde, mixed solution C with mass concentration is volume ratio of glutaraldehyde to 8% is 10-100: 1, using magnetic stirrer to stir uniformly, standing and curing, (5), the preparation obtained by curing to remove organic solvent; forming nanometer solution D. (0008).
When BSA concentration is 20 mg/mL, and mixing solution A and B at ratio is 2:1, then 2 mL of A and 1 mL of B, to have C total 3 mL, and 20 mg BSA, and mixing 3 mL of C with 8% glutaraldehyde solution at ratio 10:1, then 3 mL of C with 20 mg BSA has 0.3 mL or 300 µl of 8% glutaraldehyde solution, so 1 mg of BSA has 15 µl of 8% glutaraldehyde solution, or 18 µl of 6.5% glutaraldehyde solution.
Similarly, When BSA concentration is 20 mg/mL, and mixing solution A and B at ratio is 2:1, then 2 mL of A and 1 mL of B, to have C total 3 mL, and 20 mg BSA, and mixing 3 mL of C with 8% glutaraldehyde solution at ratio 20:1, then 3 mL of C with 20 mg BSA has 0.15 mL or 150 µl of 8% glutaraldehyde solution, so 1 mg of BSA has 7.5 µl of 8% glutaraldehyde solution, or 9.2 µl of 6.5% glutaraldehyde solution. The numbers of µl of 6.5% glutaraldehyde solution to use with 1 mg BSA can be lower or higher than 10 µl of 6.5% glutaraldehyde solution to crosslinked BSA to prepare BSA nanoparticles without or with iron particles, which also depends on one with skill in the art goals, to have more magnetic particles, more solid BSA with more crosslinking or not and other factors may play roles like long term stability, release rates, etc.
It would have been obvious to one of ordinary skill in the art before the effective filing date of the invention to prepare albumin nanoparticles loaded with magnetic nanoparticles by the steps taught by Nosrati et al. with a with vigorous mixing, constant stirring and vigorous stirring in all steps a, b, and c, with the weight ratio 10:3.75 of albumin over iron salts taught by Nosrati et al., and superparamagmetic iron-oxide nanoparticles (SPIONs), mixing at several tens of thousands of rpm by homogenizer taught by KR 101125232 B1, and to have step mixture pH adjusting to be alkaline, which is in the range of 8-12, taught by Zhang et al. (CN 103611172 B) and pH 8-10 taught by Zhang et al. (CN 104546741 A), and to apply 9.2 µl of 6.5% glutaraldehyde solution per 1 mg of BSA since together they have proven they could successfully prepare these magnetic nanoparticles.
Conclusion
5. No claim is allowed.
6. Any inquiry concerning this communication or earlier communications from the examiner should be directed to NGOC-ANH THI NGUYEN whose telephone number is (571)270-0867. The examiner can normally be reached Monday - Friday 8:00 am.
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/NGOC-ANH THI NGUYEN/Examiner, Art Unit 1615
/JESSICA WORSHAM/Primary Examiner, Art Unit 1615