CN107987060A - A kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug - Google Patents
A kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug Download PDFInfo
- Publication number
- CN107987060A CN107987060A CN201711406048.2A CN201711406048A CN107987060A CN 107987060 A CN107987060 A CN 107987060A CN 201711406048 A CN201711406048 A CN 201711406048A CN 107987060 A CN107987060 A CN 107987060A
- Authority
- CN
- China
- Prior art keywords
- formula
- nampt
- application
- antitumor drug
- cell
- 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
- C07D—HETEROCYCLIC COMPOUNDS
- C07D401/00—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom
- C07D401/14—Heterocyclic compounds containing two or more hetero rings, having nitrogen atoms as the only ring hetero atoms, at least one ring being a six-membered ring with only one nitrogen atom containing three or more hetero rings
Landscapes
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
The invention discloses a kind of sulfamide derivative formula（Ⅰ）, formula（Ⅰ）For new construction, formula is also disclosed（Ⅰ）Preparation method and its application as NAMPT inhibitor in antitumor drug.Suppressing the activity experiment of NAMPT and mtt assay measure formula（Ⅰ）Good activity is all shown in experiment to the inhibitory action of different tumour cells.Formula（Ⅰ）Structure is, wherein, R1Selected from COOH or CH2OH。
Description
Technical field
A kind of application the present invention relates to sulfamide derivative and its as NAMPT inhibitor in antitumor drug.
Background technology
Nampt（Nicotinamide phosphoribosyl transferase, NAMPT) again
Name Nampt visfatin or pre-B cell clone enhancer PBEF, regulates and controls required energy matter in mammalian cell
The level of NAD.Cancer cell has very high NAD consumption and metabolic rate, thus the rate-limiting enzyme NAMPT of NAD route of synthesis becomes
The new target drone for the treatment of of cancer, its enzyme inhibitor FK866 and CHS-828 are typical NAMPT inhibitor, field of cancer treatment into
Extensive clinical research is gone.
FK866 ((E)-N- [4- (1- benzoyl piperidine -4- bases) butyl] -3- (pyridin-3-yl) acrylamide) exists
Inducing cell apoptosis in HepG2 cells, but do not have initial action to cellular energy metabolism.FK866 can be used for treatment and involve
The disease such as cancer of Apoptosis imbalance.The prior art is it was demonstrated that the biology of FK866 interference nicotinamide adenine dinucleotide
Synthesize and apoptosis-induced cell death and without any DNA damage effect.FK866 suppresses NAMPT and exhausts the cell of NAD but do not draw
Play cytotoxicity at once, this hint, synthesize the cancer cell of NAD for anti-dependence niacinamide for, FK866 is promising medicine
Thing.In mouse mammary cancer model, FK866 also induces tumor growth delay and tumor radiosensitivity to strengthen, with NAD water
The dose dependent of flat, pH and energy state reduces.In THP-1 and K562 Leukemia Cell Lines, it has been observed that FK866 is resisted
The chemical sensitization effect of the cell death of tumour medicine 1- methyl-3-nitro -1- nitrosoguanidines (MNNG)-induction.Moved in xenogenesis
The effect of GMX1777 is evaluated in plant model and the medicine generation distribution of GMX1778 is measured and its to nicotinoyl by liquid chromatography/mass spectrometry
The effect of amine adenine-dinucleotide cellular level.
The above shows that NAMPT inhibitor has good druggability, available for preparation by suppressing niacinamide phosphoric acid
Phosphoribosynltransferase is come the medicine of disease that prevents and/or treat, it can also be used to the disease such as anti-curing cancers.New NAMPT suppresses
The exploitation of agent is also the hot spot of study of pharmacy.
The content of the invention
It is an object of the present invention to provide a kind of sulfamide derivative and its as NAMPT inhibitor in antineoplastic
Application in thing.Its structural formula（Ⅰ）For
,
Wherein, R1Selected from COOH or CH2OH。
Another object of the present invention is to provide the formula（Ⅰ）Synthetic route：
。
Another object of the present invention is to provide the synthetic method of a kind of sulfamide derivative, used in its building-up process
The condensation of sulfonic acid chloride and amine and the condensation of acyl chlorides and amine.What reaction utilized is the condensation reaction of sulfonic acid chloride and acyl chlorides and amine, instead
Answer that thing reactivity is higher, and reaction condition is relatively mild, there is very strong operability.Products therefrom also has very strong expansible
Property, the pharmacology structure with modified structure is readily synthesized on original pharmacological basis.
In condensation reaction solvent for use can be dichloromethane, chloroform, tetrahydrofuran, acetonitrile, petroleum ether,
The inert polarity such as ether, toluene or nonpolar solvent, preferably acetonitrile and tetrahydrofuran；Reaction temperature from 0-50 DEG C all can,
It is preferred that 0-10 DEG C.The present invention is optimized reaction condition and reaction scheme, either the use of solvent from economic benefit
The selection of amount, the species of reactant and dosage or reaction temperature has all carried out condition optimizing, the reaction scheme and condition of gained
It is the most route of Atom economy and the feature of environmental protection in the route that we select.
Another object of the present invention is to provide the formula（Ⅰ）Purposes, as Nampt suppress
The application of agent.
Further, the formula（Ⅰ）Application in Nampt inhibitor medicaments are prepared.
Another object of the present invention is to provide the formula（Ⅰ）Another aspect purposes, in antitumor drug should
With.
Further, the formula（Ⅰ）Application in antitumor drug is prepared.The tumor type includes lung cancer, liver
Cancer, stomach cancer, breast cancer, chronic myelogenous leukemia.
Formula according to the present invention（Ⅰ）It is real in the activity for suppressing Nampt for new construction
Test and measure formula with mtt assay（Ⅰ）Good activity is all shown in experiment to the inhibitory action of different tumour cells.
Obviously, the above according to the present invention, according to the ordinary technical knowledge and means of this area, is not departing from this hair
Under the premise of bright above-mentioned basic fundamental thought, the modification, replacement or change of other diversified forms can also be made.
Embodiment
Embodiment 1：The synthesis of intermediate 1- [(1 hydrogen benzo [d] imidazoles -1 base) sulfonyl] pyrrolidines -3- formyl chlorides
By 1 hydrogen benzo [d] imidazoles -1- chloride solutions（2.2mmol, 2.2 equivalents）0 DEG C is cooled to, by nafoxidine -3- formyls
Chlorine（2.2mmol, 2.2 equivalents）It is added in the chloride solution being vigorously stirred.Stirred at 0 DEG C after five minutes, by reaction vessel
Take out, warm to room temperature from ice bath, and stir 1.5 it is small when.Then reaction mixture is diluted with EtOAc, and passes through silica gel
Pad filtering, covers diatomite layer, it is washed with other EtOAc.Filtrate is concentrated, gained residue passes through flash chromatography
Purifying（Use the Biotage Isolera Four systems with SNAP 25g columns）, obtain 0.50g white solids 1- [(1 hydrogen benzene
And [d] imidazoles -1 base) sulfonyl] pyrrolidines -3- formyl chlorides, yield 72%）.1H-NMR (400 MHz, CDCl3) δ:
1.78(m, 1H), 2.07(m, 1H), 2.80(m, 1H), 3.56-3.71(m, 2H), 3.87(m, 1H), 4.29(m,
1H), 7.17-7.28(m, 2H), 7.52-7.59(m, 2H), 9.16(s, 1H). 13C-NMR (125 MHz, CDCl3)
δ: 31.92, 45.34, 46.45, 50.02, 113.79, 121.33, 124.89, 127.37, 129.52,
131.68, 143.81, 174.48. LC-MS(ESI, pos, ion) m/z: 314[M+H]。
Embodiment 2：The conjunction of 4- (1- ((1 hydrogen benzo [d] imidazoles -1- bases) sulfonyl) pyrrolidines -3- formamido groups) nicotinic acid
Into
It is dissolved in DCM（50mL）In 1- [(1 hydrogen benzo [d] imidazoles -1 base) sulfonyl] pyrrolidines -3- formyl chlorides
（2.2mmol）Solution is slowly added into the 4- amino-nicotinic acids for being cooled to that temperature is 0 DEG C（2.2mmol）And triethylamine（2.2mmol）
's（100mL）In solution.After adding, ice bath is removed, gained mixture is stirred at room temperature overnight, then by reaction mixture
It is poured into water, uses CH2Cl2Solution extracts.By organic extract MgSO4Dry and be concentrated under reduced pressure.Residue DCM/Et2O
1/10 grinding, be collected by filtration to obtain sepia solid 4- (1- ((1 hydrogen benzo [d] imidazoles -1- bases) sulfonyl) pyrrolidines -
3- formamido groups) nicotinic acid 0.81g, yield 89%.1H-NMR (400 MHz, CDCl3) δ: 1.74(m, 1H), 2.01
(m, 1H), 2.84(m, 1H), 3.50(m, 1H), 3.67-3.83(m, 2H), 4.51(m, 1H), 7.17-7.27
(m, 2H), 7.52-7.59(m, 2H), 7.92(s, 1H), 8.02(d, 1H), 8.46(d, 1H), 8.84(s 1H),
9.34(s, 1H). 13C-NMR (125 MHz, CDCl3) δ:31.21, 43.24, 45.14, 45.34, 113.79,
115.72, 118.85, 121.33, 124.89, 127.37, 129.52, 131.68, 141.28, 143.81,
150.95, 151.09, 169.06, 174.23. LC-MS(ESI, pos, ion) m/z: 416[M+H]。
Embodiment 3：1- ((1 hydrogen-benzo [d] imidazoles -1- bases) sulfonyl)-N- (3- (hydroxymethyl) pyridin-4-yl) pyrrole
Cough up the synthesis of -3- formamides
It is dissolved in DCM（50mL）In 1- [(1 hydrogen benzo [d] imidazoles -1 base) sulfonyl] pyrrolidines -3- formyl chlorides
（2.2mmol）Solution is slowly added into the 3- ethyoxyls -4-aminopyridine for being cooled to that temperature is 0 DEG C（2.2mmol）And triethylamine
（2.2mmol）'s（100mL）In solution.After adding, ice bath is removed, gained mixture is stirred at room temperature overnight, then will
Reaction mixture is poured into water, and uses CH2Cl2Solution extracts.By organic extract MgSO4Dry and be concentrated under reduced pressure.Residue is used
DCM/Et2O 1/10 is ground, be collected by filtration to obtain sepia solid 1- ((1 hydrogen-benzo [d] imidazoles -1- bases) sulfonyl) -
N- (3- (hydroxymethyl) pyridin-4-yl) pyrrole-3-carboxamide 0.75g, yield 85%.1H-NMR (400 MHz, CDCl3)
δ: 1.44(s, 1H), 1.76(m, 1H), 2.01(m, 1H), 2.80(m, 1H), 3.63-3.69(m, 2H), 3.81
(m, 1H), 4.41(m, 1H), 4.61(s, 2H),7.20-7.25(m, 2H), 7.52-7.59(m, 2H), 7.82(s,
1H), 7.94(d, 1H), 8.22(s, 1H), 8.52(s, 1H), 9.33(s, 1H). 13C-NMR (125 MHz,
CDCl3) δ:31.21, 43.24, 45.14, 45.34, 63.59, 113.79, 115.88, 121.33, 124.89,
127.37, 129.52, 131.68, 133.87, 143.81, 145.12, 148.41, 149.55, 174.23. LC-MS
(ESI, pos, ion) m/z: 402[M+H]。
Effect example 1：Formula（Ⅰ）Suppress the activity of Nampt
1st, the preparation of enzyme：Conversion there are into BL21 (DE3) plysS cell inoculations of recombinant plasmid (Nampt-pET28a+) in 2 × YT
In culture medium (37ug/ml chloramphenicol and 75ug/ml kanamycins), 37 DEG C of shakings are stayed overnight, with 20 times of substances after collection thalline
Long-pending fresh culture is resuspended, and 37 DEG C of cultures induce 5h to OD600 about 0.6 under the conditions of 0.3mM IPTG, 28 DEG C.Centrifugation is received
Collect thalline, and be resuspended in lysis buffer (20mM Tris-HCl pH8.0,300mM NaCl), 200W ultrasonic degradations are thin
Born of the same parents, ultrasonic 1s gaps 9s, carry out 30min altogether.By lysate in 12500rpm, 4 DEG C of centrifugation 50min Aspirate supernatants.The supernatant
Liquid is incubated 1h with Ni-NTA columns (being purchased from QIAGEN companies) in shaking on ice, then uses binding buffer (5mM successively
Imidazole, 0.5M NaCl, 20mMTris-Hcl, pH=7.5), wash buffer (40mM imidazole, 0.5M
NaCl, 20mM Tris-HCl, pH=7.5) foreign protein is washed away successively, finally with Elution buffer (200mM
Imidazole, 0.5MNaCl, 20mM Tris-Hcl, pH=7.5) elution destination protein, and carry out SDS-PAGE detections.It will wash
De- destination protein is transferred in bag filter, with Tris-HCl (20mM pH=7.5) dialysis 4~5 of sterilizing in 4 DEG C of refrigerators
It is secondary, after 20%PEG20000 concentrations, protein concentration is measured with Bradford methods.
2nd, enzyme reaction system is 25 μ l, wherein the concentration of various components is：50mM Tris-HCl (pH7.5), 0.02%
BSA, 12mM MgCl2,2mM ATP, 0.4mM PRPP, 2mM DTT, 2 μ g/mlNampt, 0.2 μM of NAM, 2%DMSO and multiple proportions
Diluted compound.The DMSO solution of the various concentrations of 0.5 μ l compounds is first added on 96 orifice plates, 20 μ l enzyme reactions is added and mixes
Solution (the enzyme reaction component in addition to substrate) is closed, after 37 DEG C of preincubate 5min, it is anti-to start to add 4.5 μ l substrate NAM solution
Should, 37 DEG C of reaction 15min heat 1min after 95 DEG C and terminate enzyme reaction.
3rd, 10 μ l, 20% acetophenones and 2M KOH are sequentially added after enzyme reaction solution after cooled on ice, on vortex mixed instrument
Mix and act on 2min after 0 DEG C, add 45 μ l88% formic acid, 70 DEG C of heating 5min, cooled on ice.
4th, using the fluorescent value at microplate reader measure excitation wavelength 382nm, launch wavelength 445nm.
5th, according to formula：E=R/ (1+ (C/IC50)S) (wherein E is enzymatic activity to+B, and C is compound concentration, R, IC50、S、B
For parameter to be fitted), enzymatic activity is fitted the curve of compound concentration in origin softwares, obtains formula（Ⅰ）'s
IC50。
1 formula of table（Ⅰ）The IC that NAMPT suppresses50Value
| IC50（nM） | |
| Embodiment 2 | 5.11±0.41 |
| Embodiment 3 | 5.24±0.30 |
Therefore deduce that, formula disclosed in this invention（Ⅰ）With the activity for suppressing Nampt, Ke Yizuo
For Nampt inhibitor, it can be used for the treatment of Nampt relevant disease.
Effect example 2：MTT（Tetrazolium bromide）Method measures formula（Ⅰ）To the inhibitory action of different tumour cells
1st, used tumour cell is：Human lung cancer cell A549, human liver cancer cells Hep G2, human liver cancer cell Bel-
7402, human gastric adenocarcinoma SGC-7901, human breast cancer cell line Bcap-37, people's chronic myelogenous leukemia cell K562.
2nd, experimental procedure
1. the cell in growth period of taking the logarithm, Trypsin Induced, 1640 cell culture fluid tune concentration of cell suspension of RPMI for 6 ×
104A/mL.Add 100 μ L of cell suspension per hole in 96 well culture plates, put 37 DEG C, 5% CO224h, cell patch are cultivated in incubator
Wall.
2. removing 1640 cell culture fluids of RPMI, 1640 cell culture fluids of RPMI of the medicine to be measured of concentration gradient are added
100 μ L, each concentration set 6 parallel holes.96 orifice plates after dosing are placed in 37 DEG C, 5% CO248h is cultivated in incubator, is inverted
The action effect of micro- Microscopic observation medicine.
Nutrient solution is discarded after the centrifugation of 3.96 orifice plates, after carefully being rushed 2 ~ 3 times with PBS, adds the RPMI containing 0.5% MTT
1640 cell culture fluid, 100 μ L, continue to cultivate 4h.
4. removing supernatant, 150 μ L dimethyl sulfoxide (DMSO)s are added per hole, low-speed oscillation 10min on shaking table is put, ties formazan
Brilliant fully dissolving.
5. the optical density in each hole is measured at enzyme-linked immunosorbent assay instrument 490nm（OD values）.
6. parallel hole OD values are represented with mean ± SD, inhibiting rate formula is calculated：[(ODControl group-ODBlank group)-(ODDrug study group-
ODBlank group)]/(ODControl group-ODBlank group)*100%。
7. using 5 data processing softwares of GraphPad Prism, by drawing amount effect curve calculation of half inhibitory concentration
（IC50）.
2 formula of table（Ⅰ）Human tumor cells IC50Value
Therefore deduce that, formula disclosed in this invention（Ⅰ）It is inhibited to six kinds of human tumor cells, cancer can be used as
The drug candidate of prevention and/or treatment carries out more deep research and development, has prompted formula disclosed in this invention（Ⅰ）It can be used for making
Standby antitumor drug.
Claims (4)
1. a kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug, its structural formula（Ⅰ）For
,
Wherein, R1Selected from COOH or CH2OH。
2. formula as claimed in claim 1（Ⅰ）, synthetic route is
。
3. formula as claimed in claim 1（Ⅰ）, the application as Nampt inhibitor.
4. formula as claimed in claim 1（Ⅰ）, the application in antitumor drug.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711406048.2A CN107987060A (en) | 2017-12-22 | 2017-12-22 | A kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201711406048.2A CN107987060A (en) | 2017-12-22 | 2017-12-22 | A kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN107987060A true CN107987060A (en) | 2018-05-04 |
Family
ID=62041469
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201711406048.2A Pending CN107987060A (en) | 2017-12-22 | 2017-12-22 | A kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN107987060A (en) |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020128232A1 (en) * | 2000-10-12 | 2002-09-12 | Henderson Scott A. | Heterocyclic angiogenesis inhibitors |
| US20060205941A1 (en) * | 2004-12-16 | 2006-09-14 | Bressi Jerome C | Histone deacetylase inhibitors |
| CN103384668A (en) * | 2010-09-03 | 2013-11-06 | 福马Tm有限责任公司 | Novel compounds and compositions for the inhibition of NAMPT |
| CN103717574A (en) * | 2011-05-04 | 2014-04-09 | 福马Tm有限责任公司 | Novel compounds and compositions for the inhibition of nampt |
| CN106916101A (en) * | 2017-02-15 | 2017-07-04 | 聚缘（上海）生物科技有限公司 | Double target spot inhibitor of NAMPT/HDAC and preparation method thereof |
-
2017
- 2017-12-22 CN CN201711406048.2A patent/CN107987060A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20020128232A1 (en) * | 2000-10-12 | 2002-09-12 | Henderson Scott A. | Heterocyclic angiogenesis inhibitors |
| US20060205941A1 (en) * | 2004-12-16 | 2006-09-14 | Bressi Jerome C | Histone deacetylase inhibitors |
| CN103384668A (en) * | 2010-09-03 | 2013-11-06 | 福马Tm有限责任公司 | Novel compounds and compositions for the inhibition of NAMPT |
| CN103717574A (en) * | 2011-05-04 | 2014-04-09 | 福马Tm有限责任公司 | Novel compounds and compositions for the inhibition of nampt |
| CN106916101A (en) * | 2017-02-15 | 2017-07-04 | 聚缘（上海）生物科技有限公司 | Double target spot inhibitor of NAMPT/HDAC and preparation method thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN106916101B (en) | NAMPT/HDAC double-target inhibitor and preparation method thereof | |
| CN101166732B (en) | Cyanopyridine derivative and use thereof as medicine | |
| Gupta et al. | Synthesis and bio-evaluation of indole-chalcone based benzopyrans as promising antiligase and antiproliferative agents | |
| AU2013216721A1 (en) | Modulators of methyl modifying enzymes, compositions and uses thereof | |
| Zhao et al. | Identification of Trypanosoma brucei leucyl-tRNA synthetase inhibitors by pharmacophore-and docking-based virtual screening and synthesis | |
| CN105143206A (en) | Pyrimidine-2,4-diamine derivatives for treatment of cancer | |
| CN110194762B (en) | Phthalazinone derivatives, preparation method and application thereof | |
| CN107406448A (en) | ((base of 4 methyl, the 2 oxypiperazin 1) methyl) 3 of N (base of 5 cyano group 4 ((2 methoxyethyl) amino) pyridine 2) 7 formoxyl 6; the particle of 4 dihydro 1,8 naphthyridines 1 (2H) formamide | |
| Wang et al. | Discovery of 7H-pyrrolo [2, 3-d] pyridine derivatives as potent FAK inhibitors: Design, synthesis, biological evaluation and molecular docking study | |
| Shan et al. | Expanding the structural diversity of diarylureas as multi-target tyrosine kinase inhibitors | |
| Jin et al. | Exploration of N-(2-aminoethyl) piperidine-4-carboxamide as a potential scaffold for development of VEGFR-2, ERK-2 and Abl-1 multikinase inhibitor | |
| CN105330653A (en) | Quinazoline derivatives | |
| CN107814788B (en) | A kind of sulfamide derivative, preparation method and its application as NAMPT inhibitor | |
| CN107987060A (en) | A kind of sulfamide derivative and its application as NAMPT inhibitor in antitumor drug | |
| CN104168958B (en) | Diaryl sulfonamide available for treatment inflammation and cancer | |
| CN107954987A (en) | A kind of sulfamide derivative and its application in antitumor drug | |
| CN107915721A (en) | A kind of sulfamide derivative and its application as Nampt inhibitor in antitumor drug | |
| CN108042538A (en) | A kind of sulfamide derivative and its application as NAMPT inhibitor | |
| CN105732624A (en) | Preparation method and application of pyrrole [4,3,2-de] quinoline-2-(1H)-ketone BRD4 protein inhibitor | |
| CN103709096B (en) | Urea type derivative used as nicotinamide ribose phosphate transferase inhibitor, as well as preparation method and application thereof | |
| CN115477639B (en) | Polysubstituted pyrimidine compound with FGFR1 as target point, and preparation method and application thereof | |
| Fang et al. | Design, synthesis and biological evaluation of E-ring modified evodiamine derivatives as novel antitumor agents | |
| CN101492410B (en) | Indirubin compounds for antineoplastic medicament | |
| CN107903246A (en) | A kind of sulfamide derivative, preparation method and its it is antitumor in application | |
| CN107954988A (en) | A kind of sulfamide derivative, preparation method and its application as NAMPT inhibitor in antitumor drug |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20180504 |
|
| WD01 | Invention patent application deemed withdrawn after publication |