CN106519017A - Purification method of tetracosactide acetate - Google Patents
Purification method of tetracosactide acetate Download PDFInfo
- Publication number
- CN106519017A CN106519017A CN201610927175.6A CN201610927175A CN106519017A CN 106519017 A CN106519017 A CN 106519017A CN 201610927175 A CN201610927175 A CN 201610927175A CN 106519017 A CN106519017 A CN 106519017A
- Authority
- CN
- China
- Prior art keywords
- tetracosactide
- fmoc
- resin
- purification
- reaction
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/435—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
- C07K14/665—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans derived from pro-opiomelanocortin, pro-enkephalin or pro-dynorphin
- C07K14/695—Corticotropin [ACTH]
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/55—Design of synthesis routes, e.g. reducing the use of auxiliary or protecting groups
Landscapes
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Organic Chemistry (AREA)
- Biochemistry (AREA)
- Gastroenterology & Hepatology (AREA)
- Zoology (AREA)
- Biophysics (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Medicinal Chemistry (AREA)
- Molecular Biology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Toxicology (AREA)
- Peptides Or Proteins (AREA)
Abstract
The invention discloses a purification method of tetracosactide acetate. The method comprises the steps of: adopting 2-Cl Resin as the starting resin carrier, conducting solid-phase synthesis, and adopting specific microwave technology reaction to connect corresponding amino acids in a tetracosactide sequence so as to obtain tetracosactide-2-Cl-Resin; carrying out cutting and sedimentation reaction on the tetracosactide-2-Cl-Resin to obtain tetracosactide crude peptide; subjecting the tetracosactide crude peptide to specific reversed-phase high-performance liquid chromatography purification, and performing freeze-drying, thus obtaining a tetracosactide pure product. The method provided by the invention lowers the liquidity pH value to improve the ion pairing ability, is beneficial to purification and purity improvement, also optimizes the purification gradient, shortens the purification time, saves the use of purification mobile phase, and reduces the production cost. Acetic acid is employed to replace trifluoroacetic acid to regulate the PH of the mobile phase, trifluoroacetic acid can be avoided in the final product, the product toxicity is lowered, and the technological process is simplified.
Description
Technical field
The invention belongs to polypeptide drugs synthesis technical field, more particularly to a kind of purification process of acetic acid tetracosactide.
Background technology
The sequence of acetic acid tetracosactide (Tetracosactide Acetate) is：
Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-Gly-Lys-Lys-Arg-
Arg-Pro-Val-Lys-Val-Tyr-Pro-OH,
Its molecular formula is C136H210N40O31S, molecular weight are 2933.46.
Tetracosactide (Tetracosactide) is a kind of 24 peptide thyroliberin analog of synthetic, the row of aminoacid
Row order is identical with 24 aminoacid of natural thyroliberin (people, cattle, pig) aminoterminal, has identical physiology with natural A CTHm
Activity.Most prominent the characteristics of is will not generally to produce antibody response, without serious side effects, is particularly well-suited to for natural
Pig thyroliberin has anaphylaxiss or invalid patient.Tetracosactide is usually used in diagnosing Adrenal cortex function insufficiency, it is also possible to
In diseases such as treatment rheumatism, rheumatic arthritis, dermatosiss and shocks.
Tetracosactide synthetic method in the market, synthesis technique is complicated, the cycle is long, and production cost is high, is unfavorable for replacing
Can gram peptide large-scale production.
The content of the invention
It is an object of the invention to provide a kind of purification process of acetic acid tetracosactide, synthesis preparation method reaction time
Short, reaction efficiency is high, low cost, the wide application prospect with production tetracosactide product.
The present invention is achieved by the following technical solutions：
A kind of purification process of acetic acid tetracosactide, comprises the following specific steps that：
(1), with 2-Cl Resin as initial resin carrier, by solid-phase synthesis, with Fmoc- protected amino acids as condensation
Raw material, is condensed corresponding aminoacid in tetracosactide sequence successively, and condensation product is reacted using specificity microwave technology, is reacted
Deprotection base after end, purification lyophilizing, obtain tetracosactide -2-Cl-Resin；
(2) tetracosactide -2-Cl-Resin is carried out cutting, sedimentation reaction, obtain the thick peptide of tetracosactide；
(3) tetracosactide thick peptide is carried out into distinctive reversed phase high-performance liquid chromatography purification, lyophilizing obtains tetracosactide pure
Product.
Further, the substitution value of step (1) the 2-Cl Resin is 0.64mmol/g.
Further, in step (1) condensation reaction with DIC/HOBt as condensing agent and activating reagent, during condensation reaction
Between be 25-30min.
Further, described in step (1), specificity microwave reaction technical step is：Condensation reaction carries out 15-20min
Afterwards, tetracosactide -2-Cl Resin are used microwave synthetic reaction 20s of specificity.
Further, the Fmoc- protected amino acids of the condensation of solid-phase synthesis described in step (1) are：Fmoc-Pro-OH、
Fmoc-Tyr(tBu)-OH、Fmoc-Val-OH、Fmoc-Arg(pbf)-OH、Fmoc-Lys(Boc)-OH、Fmoc-Gly-OH、
Fmoc-Trp(Boc)-OH、Fmoc-Phe-OH、Fmoc-His(trt)-OH、Fmoc-Glu(OtBu)-OH、Fmoc-Met-OH、
Fmoc-Ser (tBu)-OH, in the condensation reaction, aminoacid inventory is by 2-3 times of throwing resin molal quantity, the condensation reaction
Temperature is 25 DEG C.
Further, in the reaction of deprotection base described in step (1), deprotecting regent is 20% piperidines/DMF solution, is washed
The de- time is 20min.
Further, cutting sedimentation reaction step described in step (2) is：Resin cleavage twice after, cutting liquid is blown into
To in ice ether, centrifugation goes supernatant, repetitive operation to obtain the thick peptide product of tetracosactide for 3 times.
Further, the formula of the cutting liquid 100ml is：81.5ml TFA+2.5ml EDT+1ml TIS+5ml Fructus Foeniculis
Thioether+5ml H20+5g phenol, the cutting liquid volume for reactor volume 1/2.
Further, step (3) the distinctive reversed phase high-performance liquid chromatography purification step is：Purification selects C18,
5um prepares post, with A phases：0.1% acetic acid/water, B phases：0.1% acetic acid/acetonitrile is mobile phase, with B.Cone/% (2 → 65),
The Gradient program of Time.0 → 50min carries out the separating-purifying of polypeptide.
The invention has the advantages that：
By mobility pH value being reduced to improve ion pairing ability in the present invention, be conducive to purification and improve purity, and
And purification gradient is optimized, purification time is shortened, the use of purification mobile phase is saved, is reduced production cost；With acetic acid
Replace trifluoroacetic acid to adjust mobile phase PH, it is to avoid in final products, to contain trifluoroacetic acid, reduce product toxicity, simplify technique
Flow process.
Specific embodiment
Technical scheme in the embodiment of the present invention is clearly and completely described, it is clear that described embodiment is only
The a part of embodiment of the present invention, rather than the embodiment of whole.Based on the embodiment in the present invention, those of ordinary skill in the art
The all other embodiment obtained under the premise of creative work is not made, belongs to the scope of protection of the invention.
Embodiment 1
S1,2-Cl-Resin's is swelling
Weigh 2-Cl-Resin 1g to add into Peptide systhesis reactor from opening, add in Peptide systhesis reactor
Enter DCM solution submergence 2-Cl-Resin, after swelling 30min, drain solvent；
Wherein, 2-Cl-Resin substitution values are 0.64mmol/g；
The synthesis of S2, tetracosactide -2-Cl-Resin
Tetracosactide -2-Cl-Resin is：
Ser(tBu)-Tyr(tBu)-Se(tBu)r-Met-Glu(OtBu)-His(trt)-Phe-Arg(pbf)-Trp
(Boc)-Gly-Lys(Boc)-Pro-Val-Gly-Lys(Boc)-Lys(Boc)-Arg(pbf)-Arg(pbf)-Pro-Val-
Lys(Boc)-Val-Tyr(tBu)-Pro-2-Cl-Resin
The present embodiment using protected amino acid from resin start at the 1-24 corresponding protected amino acid of aminoacid and
Molecular weight is as shown in table 1 below：
Table 1
In the present invention, some conventional abbreviations have following meanings：
Fmoc：Fluorenylmethyloxycarbonyl
Boc：Tertbutyloxycarbonyl
DMF：N,N-dimethylformamide
DCM：Dichloromethane
Pbf：2,2,4,6,7- pentamethyl Dihydrobenzofuranes -5- sulphonyl
Pip：Hexahydropyridine
TFA：Trifluoracetic acid
tBu：The tert-butyl group
OtBu：The oxygen tert-butyl group
Trt：Trityl
DIC：N, N- Diisopropylcarbodiimide
DIEA：N, N- diisopropylethylamine
HOBt：I-hydroxybenzotriazole
TIS：Tri isopropyl silane
EDT：Dithioglycol
The synthetic method of S2-1, Fmoc-Pro-2-Cl-Resin
Weigh in the protected amino acid Fmoc-Pro-OH and 125mg HOBT addition 10mL centrifuge tubes of 115mg, while adding
3mL DCM solution dissolves, and in the centrifuge tube instills proper catalyst DIEA solution with dropper, activates 20s；
Reaction is added into Peptide systhesis reactor after 2-Cl-Resin of the solution after swelling with S1 is mixed
50min, reaction drain solvent after terminating, and obtain Fmoc-Pro-2-Cl-Resin；
The end socket reaction method of S2-2, Fmoc-Pro-2-Cl-Resin
Deprotection solution is prepared, the deprotection solution is the mixed solution of methanol and DIEA, and wherein, methanol is molten with DIEA
The volume ratio 1 of liquid:1,5mL deprotections solution is added into Peptide systhesis reactor with Fmoc-Pro-2-Cl-Resin and is sealed off
Reactivity site on resin, Peptide systhesis reactor is placed on the shaking table of 20-30r/min and shakes 20min；
The washing methods of S2-3, resin
The deprotection solution in Peptide systhesis reactor is drained with vacuum pump, while DMF solution washing resin is added, it is described
DMF solution adds the 1/3-1/2 that volume is Peptide systhesis reactor volume, and the decolouring that polypeptides reactive device is placed in 30r/min is shaken
60s is shaken on bed, DMF solution, repetitive operation 3 times is drained；
The removal methods of S2-4, Fmoc protection group
20% piperidines/deprotection solution removal amino protecting group FMOC is added in the Peptide systhesis reactor of S2-3, its
In, 20%-30% of the 20% piperidines/deprotection liquor capacity for Peptide systhesis reactor volume；
Peptide systhesis reactor is placed in into shake reaction 20min on the decolorization swinging table of 30r/min, after reaction terminates, is drained
Solvent；
The activation method of S2-5, protected amino acid
Weigh in 542mg protected amino acid Fmoc-Tyr (tBu)-OH and 125mg HOBT addition 10mL centrifuge tubes, while
The dissolving of 3mL DCM solution being added, proper catalyst DIEA solution being instilled with dropper in the centrifuge tube, after activation 20s, mixing is equal
It is even, obtain protected amino acid solution；
The synthetic method of S2-6, tetracosactide -2-Cl-Resin
By protected amino acid solution add into S2-5 in Peptide systhesis reactor with resin reaction, Peptide systhesis are reacted
Device is placed in 25 DEG C of isothermal vibration device and reacts 20-30min, and it is anti-that Peptide systhesis reactor is placed in microwave chemical after terminating by reaction
Microwave reaction 20s in device is answered, Fmoc-Tyr (tBu)-Pro-2-Cl-Resin after washing, is obtained；
According to above-mentioned steps connected on resin successively Fmoc-Val-OH, Fmoc-Lys (Boc)-OH, Fmoc-Val-OH,
Fmoc-Pro-OH、Fmoc-Arg(pbf)-OH、Fmoc-Arg(pbf)-OH、Fmoc-Lys(Boc)-OH、Fmoc-Lys(Boc)-
OH、Fmoc-Gly-OH、Fmoc-Val-OH、Fmoc-Pro-OH、Fmoc-Lys(Boc)-OH、Fmoc-Gly-OH、Fmoc-Trp
(Boc)-OH、Fmoc-Arg(pbf)-OH、Fmoc-Phe-OH、Fmoc-His(trt)-OH、Fmoc-Glu(OtBu)-OH、
Fmoc-Met-OH, Fmoc-Ser (tBu)-OH, Fmoc-Tyr (tBu)-OH, Fmoc-Ser (tBu)-OH aminoacid, obtain for can
Gram peptide -2-Cl-Resin；
The cutting method of S3, tetracosactide -2-Cl-Resin
Prepare 100ml cutting reagents：81.5ml TFA+2.5ml EDT+1ml TIS+5ml THIOANISOLE+5ml H20+5g
Phenol, is preserved in brown reagent bottle, tetracosactide -2-Cl-Resin is mixed with cutting reagent, and solid-to-liquid ratio is 1：10, add
React in Peptide systhesis reactor, Peptide systhesis device is placed in into shake reaction 40min on 20r/min decolorization swinging tables, cutting terminates
Add 35ml ice ether into 50ml centrifuge tubes afterwards, solution Jing cores in Peptide systhesis reactor are filled in ice ether, are covered
Upper centrifuge tube lid, shakes centrifuge tube up and down, and mix homogeneously is put into centrifuge tube in centrifuge, 3000r/min centrifugation 3min,
Supernatant is abandoned, repetitive operation obtains the thick peptide of emulsion state tetracosactide for 3 times；
The thick peptide of the tetracosactide is：
Ser-Tyr-Ser-Met-Glu-His-Phe-Arg-Trp-Gly-Lys-Pro-Val-Gly-Lys-Lys-Arg-
Arg-Pro-Val-Lys-Val-Tyr-Pro；
The purification process of the thick peptide of S4, tetracosactide
Tetracosactide thick peptide is dissolved with 15% acetonitrile/water, the inverted high performance liquid chromatography purification of thick peptide solution is obtained
Acetic acid tetracosactide sterling, wherein, purification condition is, chromatographic column is C18, and 5 μm prepare post, mobile phase be A (0.1% acetic acid/
Water)/B (0.1% acetic acid/acetonitrile), Gradient program is
Target acetic acid tetracosactide solution is collected, is weighed after lyophilization, calculating acetic acid tetracosactide sterling yield is
82.3%, purity is 99.9%.
Above content is only citing made for the present invention and explanation, and affiliated those skilled in the art are to being retouched
The specific embodiment stated is made various modifications or supplements or substituted using similar mode, without departing from invention or super
More scope defined in the claims, all should belong to protection scope of the present invention.
Claims (9)
1. a kind of purification process of acetic acid tetracosactide, it is characterised in that comprise the following specific steps that：
(1) it is, with 2-Cl Resin as initial resin carrier, by solid-phase synthesis, former by condensation of Fmoc- protected amino acids
Expect, successively aminoacid corresponding in condensation tetracosactide sequence, condensation product is reacted using specificity microwave technology, reaction knot
Deprotection base after beam, purification lyophilizing, obtain tetracosactide -2-Cl-Resin；
(2) tetracosactide -2-Cl-Resin is carried out cutting, sedimentation reaction, obtain the thick peptide of tetracosactide；
(3) tetracosactide thick peptide is carried out into distinctive reversed phase high-performance liquid chromatography purification, lyophilizing obtains tetracosactide sterling.
2. the purification process of a kind of acetic acid tetracosactide according to claim 1, it is characterised in that：Step (1) 2-
The substitution value of Cl Resin is 0.64mmol/g.
3. the purification process of a kind of acetic acid tetracosactide according to claim 1, it is characterised in that：In step (1)
With DIC/HOBt as condensing agent and activating reagent, condensation reaction time is 25-30min to condensation reaction.
4. the purification process of a kind of acetic acid tetracosactide according to claim 1, it is characterised in that：Described in step (1)
Specificity microwave reaction technical step is：After condensation reaction carries out 15-20min, by tetracosactide -2-Cl Resin using special
Microwave synthetic reaction 20s of property.
5. a kind of purification process of the acetic acid tetracosactide according to claim 1 or 3, it is characterised in that institute in step (1)
State solid-phase synthesis condensation Fmoc- protected amino acids be：Fmoc-Pro-OH、Fmoc-Tyr(tBu)-OH、Fmoc-Val-OH、
Fmoc-Arg(pbf)-OH、Fmoc-Lys(Boc)-OH、Fmoc-Gly-OH、Fmoc-Trp(Boc)-OH、Fmoc-Phe-OH、
Fmoc-His (trt)-OH, Fmoc-Glu (OtBu)-OH, Fmoc-Met-OH, Fmoc-Ser (tBu)-OH, the condensation reaction
Middle aminoacid inventory is 25 DEG C by 2-3 times of throwing resin molal quantity, the setting-up point.
6. the high-efficiency synthesis method of a kind of bivalirudin according to claim 1, it is characterised in that：Described in step (1)
In the reaction of deprotection base, deprotecting regent is 20% piperidines/DMF solution, and elution time is 20min.
7. the high-efficiency synthesis method of a kind of bivalirudin according to claim 1, it is characterised in that：Described in step (2)
Cutting sedimentation reaction step is：Resin cleavage twice after, cutting liquid is blown in ice ether, supernatant is removed in centrifugation, repeat
Operation obtains the thick peptide product of tetracosactide 3 times.
8. the high-efficiency synthesis method of a kind of bivalirudin according to claim 7, it is characterised in that：The cutting liquid
The formula of 100ml is：81.5ml TFA+2.5ml EDT+1ml TIS+5ml THIOANISOLE+5mlH20+5g phenol, the cutting
Liquid product is the 1/2 of reactor volume.
9. the high-efficiency synthesis method of a kind of bivalirudin according to claim 1, it is characterised in that：Step (3) spy
Some reversed phase high-performance liquid chromatography purification steps are：Purification selects C18,5um to prepare post, with A phases：0.1% acetic acid/water, B
Phase：0.1% acetic acid/acetonitrile is mobile phase, and with B.Cone/% (2 → 65), the Gradient program of Time.0 → 50min carries out polypeptide
Separating-purifying.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610927175.6A CN106519017A (en) | 2016-10-24 | 2016-10-24 | Purification method of tetracosactide acetate |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610927175.6A CN106519017A (en) | 2016-10-24 | 2016-10-24 | Purification method of tetracosactide acetate |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN106519017A true CN106519017A (en) | 2017-03-22 |
Family
ID=58293110
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610927175.6A Pending CN106519017A (en) | 2016-10-24 | 2016-10-24 | Purification method of tetracosactide acetate |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN106519017A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108070019A (en) * | 2017-12-12 | 2018-05-25 | 安徽省国平药业有限公司 | A kind of synthetic method of polypeptide chelate metal ion |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1486104A (en) * | 2002-08-21 | 2004-03-31 | 株式会社Ntt都科摩 | Radio communication method ,radio base station and radio terminal |
| CN103159850A (en) * | 2013-03-26 | 2013-06-19 | 深圳翰宇药业股份有限公司 | Preparation method of tetracosactide |
-
2016
- 2016-10-24 CN CN201610927175.6A patent/CN106519017A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1486104A (en) * | 2002-08-21 | 2004-03-31 | 株式会社Ntt都科摩 | Radio communication method ,radio base station and radio terminal |
| CN103159850A (en) * | 2013-03-26 | 2013-06-19 | 深圳翰宇药业股份有限公司 | Preparation method of tetracosactide |
Non-Patent Citations (1)
| Title |
|---|
| 邵明香: "47种肽的微波辅助固相合成工艺研究", 《现代药物与临床》 * |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108070019A (en) * | 2017-12-12 | 2018-05-25 | 安徽省国平药业有限公司 | A kind of synthetic method of polypeptide chelate metal ion |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108059666B (en) | A kind of method that solid-liquid combination prepares Suo Malu peptides | |
| CN106478806B (en) | A kind of solid phase synthesis process of Suo Malu peptide | |
| CN113880936B (en) | Solid-phase synthesis method of abamectin | |
| CN102647996A (en) | Relaxin peptide synthesis | |
| CN106589069B (en) | A kind of preparation method of oxytocin | |
| CN103304659A (en) | Solid-phase preparation method of liraglutide | |
| CN106397573A (en) | Solid-phase synthesis method of liraglutide | |
| CN106518966A (en) | Synthetic method of RGD cyclopeptide | |
| CN107056894A (en) | A kind of method of fragment method synthesis in solid state ganirelix acetate | |
| CN106478805A (en) | A kind of preparation method of GLP-1 derivant | |
| CN109021092A (en) | A kind of synthetic method of Suo Malu peptide | |
| CN108059667B (en) | A kind of solid phase synthesis process of Lanreotide | |
| CN106519017A (en) | Purification method of tetracosactide acetate | |
| CN106478778A (en) | A kind of solid phase synthesis process of MALDI-PSD | |
| CN108070030B (en) | Preparation method of loxenapeptide and analogue thereof | |
| CN104761619A (en) | Desmopressin acetate solid phase preparation technology | |
| CN114230653B (en) | Preparation method of chlorotoxin | |
| CN108976296A (en) | A kind of synthetic method of Liraglutide | |
| CN112321699B (en) | Synthesis method of semaglutide | |
| CN106478813A (en) | A kind of high-efficiency synthesis method of bivalirudin | |
| CN106518979B (en) | A kind of synthesis preparation method of Leuprorelin | |
| CN112521483A (en) | Preparation method of ularitide | |
| CN107417786B (en) | Preparation method of thymosin alpha 1 | |
| CN112279891A (en) | Improved solid-phase synthesis method of triptorelin | |
| CN103992401B (en) | Method for preparing exenatide |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20170322 |
|
| RJ01 | Rejection of invention patent application after publication |