Title of Invention

NOVEL INTERMEDIATES USEFUL FOR THE PREPARATION OF COENZYMES, PROCESS FOR THE PREPARATION OF THE SAID NOVEL INTERMEDIATES, AND AN IMPROVED PROCESS FOR THE PREPARATION OF COENZYMES

Abstract The present invention relates to novel intermediates for the preparation of coenzymes, processes for the preparation of the intermediates and an improved process for the preparation of Coenzymes. The present invention particularly relates to an improved process for the preparation of Coenzyme Q, more particularly for Coenzyme Q 9 and Coenzyme Q10. Still more particularly this invention relates to regio and stereo controlled process for the preparation of Coenzyme Q9 and Coenzyme Q10 of the formula 1n. where n =9 (Coenzyme C0Q9), and where n=10(Coenzyme C0Q10).
Full Text FORM-2
THE PATENTS ACT 1970 (Act 39 of 70)
COMPLETE SPECIFICATION
(Section 10)
NOVEL INTERMEDIATES USEFUL FOR THE PREPARATION OF COENZYMES, PROCESS FOR THE PREPARATION OF THE SAID NOVEL INTERMEDIATES AND AN IMPROVED PROCESS FOR THE PREPARATION OF COENZYMES
NICHOLAS PIRAMAL INDIA LIMITED, a company registered under the Indian Companies Act 1956 having its registered office located at Nicholas Piramal Tower, Peninsula Corporate Park, Ganpatrao Kadam Marg, Lower Parel, Mumbai 400 013, Maharashtra, India
The following specification particularly describes the nature of this invention and the manner in which it is to be performed



Introduction
The present invention relates to novel intermediates for the preparation of coenzymes, processes for the preparation of the intermediates and an improved process for the preparation of Coenzymes. The present invention particularly relates to an improved process for the preparation of Coenzyme Q, more particularly for Conenzyme Q 9 and Coenzyme Q10. Still more particularly this invention relates to regio and stereo controlled process for the preparation of Coenzyme Q9 and Coenzyme Q10 of the formula 1„.




Where n =9 (Coenzyme CoQ 9), and where n =1o.( Coenzyme C0Q10)
In the description given below the Coenzyme CoQ 9 is referred to as having the formula 1$ and Coenzyme CoQioas haying the formula l10
These coenzymes belong to the class of ubiquinones that occur in all aerobic organisms from bacteria to plants and animals - the name ubiquinone suggests its ubiquitous occurrence. They are involved in mitochondrial processes such as respiration and act as antioxidants.
The present invention also provides novel Grignard reagent which is useful for he preparation of above mentioned coenzymes and a process for its preparation
The coenzyme Q in human has 10 isoprenoid units, and termed as CoQ 10. CoQ 10 is present in virtually in every cell in the human body and is known as the "miracle nutrient", and plays a vital role in maintaining human health and vigor, maintenance of heart muscle strength, enhancement of the immune system, quenching of free radical in the battle against ageing to name a few ( "The miracle nutrient coenzyme" Elsvier/ North - Holland Biomedical Press, New York, 1986; " Coenzyme Q: Bioechemistry, Bioenergetics, and clinical Applications of Ubiquinone" Wiley, New
2

York, 1985; " Coenzyme Q, Molecular Mechanism in Health and Disease" CRC press).
Prior Art
As depicted above Coenzyme Q9 and Coenzyme Q10 the formula 1_ have 2,3 dimethoxy 1,4 benzoquinone nucleus as a head group with a side chain of n isoprene units. The poly prenyl side chain in Coenzyme Q has all - trans configuration. One of the methods of synthesis of these Coenzymes is coupling of the "benzoquinone nucleus" with the "polyprenyl side chain " of solanesol of the formula 3a 9, where x = -OH and decaprenol of the formula 3a1o where x = -OH. with retention of its original double bond geometry.

Various methods for introducing polyprenyl side chain into quinone nucleus, to prepare Coenzymes are found in literature. These methods involve functionalisation of the two coupling partners, the "quinone nucleus" and the "polyprenyl chain".
3
Method 1 Polyprenyl alcohol and hydroquinone using Zinc Chloride as catalyst; reported in Huanxue Yu Nianhe(2002), 6 267(2002) which is shown in the Scheme 1 given below


Decaprenol of the formula 3a 10 (1.8 g) dissolved in ether is treated with 2,3 dimethoxy-5-methyl benzohydroquinone of the formula (4), zinc Chloride (anhydrous, 0.28 g), glacial acetic acid (0.02 ml) and stirred for 2 hours under nitrogen atmosphere. Ferric chloride solution is added to the above reaction mixture, stirred for ten minutes. The ethereal layer is then separated dried and evaporated to give 2.2 g of crude CoQ1owhich is purified by column chromatography to give 0.56 g of the pure CoQ1o of the formula 1 with an overall yield of 20% (mp 45-46 ° C, Lit. mp 48-50 °C).
Low melting point obtained indicates the presence of cis isomer and thereby making the process not stereoselective. The yield is also too low for commercialization of the process.
Method 2 By making n -Allyl Nickel bromide complex and protected quinone nucleus; reported in Bull. Chem. Soc. Jpn 47,3098(1974), US 3,896,153(1975) which is shown in scheme 2

Nickel tetracarbonyl 4.5g (15% solution in Benzene) is treated with decaprenyl bromide of the formula (3b10) 10.0 g (15% solution in Benzene)at 50°C for 4-4.5 hrs. The solution is cooled to below 10°C and the benzene and excess nickel carbonyl is removed under reduced pressure.. Decapernyl nickel bromide of the formula 5 thus formed is then reacted with 6-bromo-2,3 dimethoxy -5-methyl 1,4 hydroquinone diacetate of the formula 6 in 30 ml of hexamethyl phosphoramide at 75° C for 7 hours yielding 2.2g of condensed product of the formula 7 with 40% yield. The condensed
4

product of the formula 7 (0.8 g) is added to a suspension of lithium aluminium hydride in 20 ml of dry ether and refluxed for 24 hours. The excess lithium aluminium hydride is decomposed and the product hydroquinone is extracted in ether. The hydroquinone is oxidized with aqueous ferric chloride at room temperature for 3 hour to give the final product CoQ1o which is further purified by column chromatography to yield CoQ1othe formula 1 with mp 20-22 ° C(Lit. mp 48 -50 ° C) with 69% yield.
Author attributes the low melting point to the presence of cis isomer. The process is therefore not setereoselective . Further, the nickel tetracarbonyl used in the process is highly flammable, has the risk of explosion and highly toxic chemical, and can not be used industrially. The overall yield of the process is only 27.6%. The process is therefore not suitable for industry.
Method 3 From allyl - stannyl and unprotected quinone using borontrifluoride etherate; reported in J. Org. Chem. 45,4077(1980), Chemistry Letters 885(1979 as shown in scheme 3 ).

Trimethylstannyl lithium in tetrahydrofuran is slowly added to decaprenyl bromide of the formula 3b10 at -78 C to - 60 C and the reaction mixture is allowed to warm to room temperature. The reaction mixture is quenched in brine and the organic layer evaporated to form trimethyl decaprenyl stannanes of the formula (9). The stannyl reagent (0.42 mmol) in a mixture of methylene dichloride (25 ml) and isooctane(l ml) is added to 2,3 dimethoxy-5-methylbenzoquinone(0.111g, 0.61 mmol) and borontrifluoride etherate (2.6 mmol) in a mixture of methylene chloride(25 ml) and isooctane(l ml) at - 50 C and the reaction mixture is maintained at the same temperature for 2 hours. The resulting product is isolated and chromatographed on silica gel to afford the starting quinone(70 mg) and CoQ1o of the formula l10 ( 189 )mg (86% trans).
5

The method forms 14% cis isomer and therefore far from stereo selective. The reaction does not go to completion and results in poor yield and not suitable for industry.
Method 4 From polyprenyl alcohol and quinone nucleus with silica - alumina as catalyst reported in US 3,998,858(1976) as shown in scheme 4

2,3 dimethoxy-5 -methyl-1,4-benzohydroquinone of the formula 4, l lg is reacted with boric acid 3.6g in toluene and water removed azeotropically. The residue is treated with silica-alumina(17 g) and a solution of decaprenol ( 14 g in 10 ml hexane, 94% purity) and stirred for 1 hour at 30 ° C. The adsorbent is removed and the filtrate is washed with water, and concentrated, and extracted in ether. The ethereal extract is treated with silver oxide 6 g and allowed to stand overnight. The reaction mixture is filtered and concentrated to form 16.3 g of crude CoQ1o, which is purified by column chromatography, followed by crystallization with acetone to give 8.5g of CoQ1o of the formula l1o
The melting point value indicates that process may form a stereoselective process using a simple technique of silica-alumina. However the ratio of silica and alumina to be used and also the respective grades would be critical for the reaction and is not mentioned. We tried various grades of silca - alumina and found that the reaction does not proceed.
Method 5 Polyprenyl alcohol and quinone nucleus reported in Chemistry Letters 1597(1988), as shown in scheme 5


Isodecaprenol compound of the formula 10(38.8 g, 72% purity) is reacted with 2,3 dimethoxy 5 methyl 1,4 benzohydroquinone compound of formula 4(75.1 g) in the presence of borontrifluoride etherate in hexane and nitromethane at 43 C . The reaction mixture is quenched in aqueous medium and the nitromethane and the hexane layer is separated. The hexane layer is oxidized with ferric chloride hexahydrate in isopropanol at room temperature . The crude CoQ1o of the formula l1o is obtained in 51% yield with 8% Z isomer
The process forms 8% cis isomer and therefore not stereo selective. Boron trifluoride etherate is a corrosive chemical and not useful for commercialisation.
Thus literature does not provide a stereoselective process for coupling of the benzoquinone with the polyprenyl side chain for the preparation of Coenzymes Q , namely CoQ 9 and CoQ 10. As shown in the coupling reactions mentioned above , 8% - 15% of ds isomer is formed.
We observed that purification of such a mixture to get the desired all- trans isomer of CoQ 9 and CoQ 10 with less than 1% ds, results in 25-30%) purification loss. This would decrease the overall yield of production of these coenzymes mainly CoQ 9 and CoQ 10 , thereby making the commercial process of making the Coenzyne Q9 or Coenzyme Q 10 cost ineffective.
Scope of clinical application of coenzymes specially CoQ 10 is becoming wider with its increasing broadband use Therefore if a cost effective process is developed for the preparation of COQ1oit will greatly help in making this coenzyme easily and at affordable prices
7

Preparation of coenzymes CoQ1o where n represents the number of isoprenyl units, namely CoQ 9 or CoQ1o, by the coupling of the two key units viz. the "benzoquinone nucleus" and the "polyprenyl side chain" should be a straightforward route. However as discussed in prior art, the attempts with such coupling, results in isomerisation of the polyprenyl chain and the geometrical configuration of the chain is not retained. Therefore, the focus should be on the "stereoselective" coupling reaction of the "benzoquinone nucleus" with the corresponding "polyprenyl side chain" to obtain CoQn where n represents the number of isoprenyl units. Such a condensation would enhance the cost effectiveness of the preparation of these coenzymes mainly Q9 or Q 10.
With our experience in this area, we observed that a simple , straightforward , stereo selective process for the preparation of coenzyme CoQ 9 or CoQ 10 of the formulae 1 9 and 1 10 respectively can be developed, by Grignard coupling of the benzoquinone nucleus and the polyprenyl side chain . For such a coupling the " benzoquinone nucleus" has to be converted to the required Grignard reagent with suitable protecting groups. The protecting groups used in literature for making Grignard reagent of the "benzoquinone nucleus" are methoxyethoxymethyl and methyl of the formula 11a & lib.

Literature method for making Grignard reagent compound of formula 11a from the compound of the formula 2 as reported in J. Org.Chem.37 1889(1972), US 4,270,003(1981), Synthesis (1981) 469-471(1982) comprises the methods as depicted in Scheme 6a and Scheme -6b.


In the method described in the Scheme 6a , 2,3 dimethoxy -5-methyl 1,4 benzoquinone compound of the formula 2 is brominated to form compound of formula 12 . The bromination is effected using bromine in carbon tetrachloride and the product of the formula 12 is isolated by washing with ethanol and recrystallizing from petroleum ether, in 74% yield. The compound of the formula 12, is reduced employing aqueous sodium hydrosulphite solution in presence of methanol to get the compound of the formula 13. The compound of the formula ,13 is finally converted to compound of the formula 14a by alkylation .The alkylation is carried out in presence of 50% sodium hydride in mineral oil which is (106 g) added in small portions to a stirred solution of 6 bromo 2,3 dimethoxy -5-methyl hydroquinone compound of formula (262.9 g) in 4 litres of N,N dimethyl formamide at -20 ° C. Chloromethyl
2-methoxyethyl ether(273 g) is added dropwise over a 2 h period and the mixture is allowed to warm to room temperature. Excess sodium hydride is destroyed with ethanol and the reaction mixture quenched in water. The ethereal layer containing the extracted product is concentrated and the residue purified by column to obtain the compound of formula 14a in 91% yield. The compound of the formula 14a is converted to the compound of the formula 11a , by reacting with Magnesium in presence of tetrahydrofuran.
Yield of brominating 3, 4 dimethoxy -5-methyl 1,4 benzoquinone, is only 74% which is low for such a simple reaction. The solvent used is toxic and not suitable for scale up. We observed that reduction using aqueous sodium hydrosulphite solution gives yield of the compound of the formula 13 in not more than 40% and therefore not suitable for the industrial production . Further we observed that bromination followed by reduction of the benzoquinone to obtain compound of formula .13, results in low purity of not more than 76%.
9

The alkylation process uses N,N dimethyl formamide as a solvent and in large excess , 15 times the weight of the bromo compound of the formula (13). N, N dimethyl formamide is a costly solvent and such large excess is not suitable for industry. Sodium hydride used as a base is hazardous and is always present in suspension in oil. The oil also gets extracted in the solvent in which the product compound of formula 14a gets extracted. Thus the process is not compatible to the industry.
Another method of making 2,3 dimethoxy 5-bromo 6-methyl 1,4 hydroquinone is shown in Scheme 6 b

In this method, bromination is carried out of 2,3 dimethoxy 1,4 hydroquinone of formula 4 in chloroform at 5 C, and the product isolated from chloroform in quantitative yield.
We observed that bromination at 5 °C leads to incompletion of reaction and isolation of product from chloroform results in yield.less than75%
The Grignard reagent of formula l1b is prepared as given in scheme 6C





2 12 13 14b l1b
Scheme - 6C
10

In the process depicted in Scheme 6C, 2,3 dimethoxy 5 methyl benzoquinone of the formula 2 is brominated in room temperature in carbon tetrachloride in 75% yield, reduced with Zinc and acetic acid with 80% yield and methylated with dimethyl sulphate to get the compound of the formula 14b in 62% yield. The compound of the formula 14 b is converted to compound of the formula 1 lb. Yield at each stage of the process is not substantial for mass scale production.
We observed that the above process of reduction with zinc and acetic acid, and methylation after bromination results in purity of compound of formula 14b in not more than 76%.
Therefore, to provide a simple, straightforward, stereoselective process for the preparation of coenzyme CoQ 9 or C0Q10 by coupling of the benzoquinone nucleus with polyprenyl side chain using Grignard reaction for the preparation of CoQ 9 & CoQ 10, it is essential to improve the process of preparing the Grignard reagents of the formula 11 a and 11 b , which would avoid the drawbacks of the hither to known processes exemplified above
While developing the improved process for the preparation of the Grignard reagents of the formulae 11a and 11 b , we developed a new Grignard reagent of the formula 18


18
Objective of the invention
The main objective of the present invention is to provide an improved process for the stereoselective preparation of the Coenzymes namely, CoQ 9 and CoQ 10 of the formulae 1_9 and l_1o respectively as given above
11

Another objective of the present invention is to provide an improved process for the preparation of the coenzymes, namely, CoQ 9 and CoQ 1o of the formulae 1 9 and 1 10 respectively, which is simple, cost effective and commercially viable.
Still another objective of the present invention is to provide an improved process for the preparation of the coenzymes Q, namely, CoQ 9 and C0Q10 of the formulae 1 9 and 1 10 respectively with high yield (50-56 %) and purity 98%
Still another objective of the present invention is to provide a novel Grignard reagent of the formula 18( shown above) useful for preparing the coenzymes namely, CoQ 9 and CoQ10of the formulae l9and 110 respectively
Yet another objective of the present invention is to provide a process for the preparation of novel Grignard reagent of the formula 18 useful for the preparation of the coenzymes, namely,CoQ 9 and C0Q10 of the formulae I9 and 110 respectively by coupling of "benzoquinone nucleus" with "polyprenyl side chain".
Yet another objective of the present invention is to provide an improved process for the preparation of Grignard reagents of the formula 11a and l1b (shown above)useful for preparing CoQ 9 and C0Q10 of the formulae 1_9 and 1 10 respectively by coupling of "benzoquinone nucleus" with "polyprenyl side chain".
Yet another objective of the present invention is to provide an improved process for the preparation of coenzymes I9 and 110 by sterospecific coupling of the polyprenyl side chain of formula 3b_9 or 3b 1o_with the Grignard reagents of the formula 11a or l1b or 18.
Still another objective of the present invention is to provide a novel intermediate of the formula la 9 useful for preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a process for the preparation of novel intermediate of the formula la 9 useful for preparing the coenzyme CoQ 9
12

Still another objective of the present invention is to provide a novel intermediate of the formula lb 9 useful for preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a process for the preparation of novel intermediate of the formula lb 9 useful for the preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a novel intermediate of the formula 19_ useful for preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a process for the preparation of novel intermediate of the formula 19_ useful for the preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a novel intermediate of the formula 20_ useful for preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a process for the preparation of novel intermediate of the formula 20 useful for preparing the coenzyme CoQ 9
Still another objective of the present invention is to provide a novel intermediate of the formula 21_ useful for preparing the coenzyme CoQ 10
Still another objective of the present invention is to provide a process for the preparation of novel intermediate of the formula 21 useful for preparing the coenzyme CoQ 10
The process for the preparation of the new Grignard Reagent of the formula 18, according to the present invention is shown in the Scheme 7
13


The improved process for the preparation of the Grignard reagent of the formula 11 a according to the invention is shown in Scheme 8

The improved process for the preparation of the Grignard reagent of the formula lib according to the present invention is shown in Scheme 9

Scheme 9
The improved processes for the preparation of the coenzymes, namely, CoQ 9 and C0Q10 of the formulae 19 and 110respectively, according to the invention, is shown in the Schemes 10-15 given below
14

Process for the preparation coenzyme, CoQ9 of the formula 1 9 using dimethoxyethoxymethyl as protecting groups in Grignard reagent of the formula 11a, through novel compound of the formula 1 a 9

Process for the preparation of coenzyme CoQ 9of the formula 1 9 using methyl as protecting groups in Grignard reagent of the formula lib through novel compound of the formula 1b9

15


16
Process for the preparation of compound of the formula 19 using methoxyethoxymethyl and methyl as protecting groups in novel Grignard reagent of formula 18, through novel compounds of the formula 19 and formula 20 respectively


Process for the preparation of coenzyme C0Q10 using dimethoxyethoxymethyl as protecting groups in Grignard reagent of the formula 11a through compound of the formula la1o

17
Process for the preparation of coenzyme, C0Q10 using methyl as protecting groups in Grignard reagent of the formula l1b through compound of formula lb10








Scheme -14a
Process for the Preparation of coenzyme CoQ 1o using methoxyethoxymethyl and methyl as protecting groups in novel Grignard reagent of the formula 18 through the novel compounds of the formula 21. and 22

According to the present invention there is provided a process for the preparation of novel Grignard reagent of the Formula 18
18


which comprises,
by known method, to obtain compound of formula 16,
(i) Brominating the compound of the formula 15


19
(ii) Alkylating the compound of the formula 16 obtained in step (ii ) with methoxyethoxymethyl chloride in the presence of a base, an alkali metal alkoxide or metal hydride,to obtain2,3-dimethoxy-5-methyl-6-bromo hydroquinonel,4 dimethoxyethoxymethyl ether compound of formula 17


(iii)Reacting the compound of the formula 17 obtained in step (ii) with Magnesium in presence of ether at 0-65 ° C, in presence of iodine and dibromoethane, to obtain the novel Grignard reagent of the formula 18
(iv) cooling the resulting reaction mixture to room temperature , filtering to get the novel Grignard reagent in solution .
The above process is shown in the reaction Scheme 7 given above
The compound of formula 15 can be prepared by any known methods. However in our copending application no. we have described and claimed an improved process for the preparation of compound of formula 15
Synthesis of this novel Grignard reagent is most economical as it can be made from the compound of formula 15, unlike the known Grignard reagents of formula Ha and l1b that are made from 2,3 dimethoxy-5-methyl 1,4 benzoquinone (CoQ o), thereby having more number of steps in their preparation. Presence of only one protecting group of methoxy- ethoxymethyl in compound of formula 18, reduces the requirement of the reagent methoxyethoxyethyl ether as compared to that required in dimethoxyethoxymethyl ether in 11a and make it more cost effective. At the same time cleaving of the protecting group of the formula 18 employed in the present invention results in the formation of the moiety "2,3,4 trimethoxy 6-methyl phenol" that can be easily oxidised with an inexpesive chemical like ferric chloride unlike eerie ammonium nitrate an expensive oxidising agent required for methyl protection when compound of formula l1b is used.
The novel Grignard reagent of formula 18 has also been used in the synthesis of CoQ io in our copending patent application no
20
Accordingly, the present invention provides an improved process for the preparation of Grignard reagent of the formula 11a ,


which comprises
(i)Reducing 2,3 dimethoxy -5 -methyl 1,4 benzoquinone (CoQ1o) of the formula 2

With sodium hydrosulphite, in alkaline medium, in biphase, in the presence of a water
immiscible organic solvent, and
ii separating the organic phase,
iii evaporating the oraganic phase obtained in step ii to obtain a concentrated
residue of compound of formula 4

iv Adding the concentrated residue obtained in step (iii) to a hydrocarbon solvent to
precipitateout the soild of the compound of formula 4and filtering out the solid
v Brominating the resulting compound of the formula 4 with bromine in
chlorinated hydrocarbon at 0-25 ° C ,
vi Quenching the resultant reaction mixture in step (v) in aqueous medium to
obtain aqueous and organic phase and separating the oraganic phase,
vii Evaporating the organic phase obtained in step (vi) to obtain a concentrated
21
residue of 2,3 dimethoxy -5 -methyl -6-bromo 1,4 hydroquinone of the formula 13


viii Adding the concentrated residue obtained in step vii to a hydrocarbon solvent
to precipitate out the soild of the compound of formula 13 and filtering out the solid
ix Alkylating the 2,3 dimethoxy -5 -methyl -6-bromo 1,4 hydroquinone of the
formula 13 obtained in step (viii) with methoxyethoxymethyl chloride in the presence of a base, an alkali metal alkoxide or metal hydride, to obtain 2,3-dimethoxy-5-methyl-6-bromo hydroquinone 1,4 dimethoxyethoxymethyl ether compound of formula 14a,

and
x Reacting the compound of the formula 14a obtained in step (ix ) with
Magnesium in presence of ether, iodine and dibromoethane, at a temperature in the
range of 0-65 C, to obtain the Grignard reagent of the formula 11 a
The above mentioned process is illustrated in the reaction Scheme 8 shown above In the above mentioned process the improvements were effected by carrying out the reduction in step (i) to obtain compound of formula 4_using aqueous hydrosulphite in biphase, in alkaline medium in the presence of a water immiscible organic solvent. The hydroquinone of formula 4 was isolated by precipitating out the soild in presence of a hydrocarbon solvent In the prior art process this reduction is effected in homogeneous phase using water miscible solvent. The modified process according to the present invention increases the yield of the reduced product of the formula 4 substantially (to about 96 % as compared to about 50% as per the prior art process). According to the improved process of the present invention , the brominated product compound of formula 13 was isolated by precipitating out the soild in presence of a hydrocarbon solvent. The process described above increases the yield of the brominated compound (to about 96 % as compared to 75 % as per the prior art process)
22

In the modified process of the present invention the alkylation is carried out in the
presence of a base sodium hydride in an inexpensive hydrocarbon solvent, or
nonhazadrous sodium alokoxide, in an inexpensive solvent alcohol. Thereby making
the process economical as compared to prior art where sodium hydride is used in
presence of N,N dimethyl formamide which is an expensive solvent
By reacting the bromo compound with magnesium in the presence of ether like
diethylether, diisopropyl ether, tetrahydrofuran, at a temperature in the range of 0 -
65 ° C , resulting in the production of Grignard reagent of the formula 11a having
92 % purity.
All the above said modifications effected to the known process of preparing
compounds of the formulae 11a , impart novelty to the process defined above
According to another feature of the present invention there is provided an improved process for the preparation of Grignard reagent of the Formula l1b ,

which comprises,
(i) Reducing 2,3 dimethoxy -5 -methyl 1,4 benzoquinone (CoQ1o ) of the

formula 2
With sodium hydrosulphite, in alkaline medium, in biphase, in the presence of a water
immiscible organic solvent, and
ii separating the organic phase,
iii evaporating the oraganic phase obtained in step ii to obtain a concentrated
residue of compound of formula 4
23



iv Adding the concentrated residue obtained in step iii to a hydrocarbon solvent to precipitate out the soild of the compound of formula 4and filtering out the solid v.Alkylating the compound of the formula 4_, with alkyl sulphate by known method to obtain 2,3,4,5 tetramethoxy toluene compound of formula 4b

vi Brominating the resulting compound of the formula 4b with bromine in
chlorinated hydrocarbon at 0-25 ° C ,
vii Quenching the resultant reaction mixture in step (vi) in aqueous medium to obtain
aqueous and organic phase and separating the oraganic phase,
viii Evaporating the organic phase obtained in step vii to obtain a concentrated
residue of 2,3,4,5 tetramethoxy 6-bromo toluene of the formula 14b

ix Adding the concentrated residue obtained in step viii to a hydrocarbon solvent to precipitate out the soild of the compound of formula 14b and filtering out the solid

x Reacting the compound of the formula 14b obtained in step (iii) with Magnesium in presence of ether, iodine and dibromoethane at 0-65 C, to obtain the Grignard reagent of the formula l1b

This process has been shown in Scheme no 9 given above
In the above mentioned process the improvements were effected by carrying out the reduction in step (i) to obtain compound of formula 4_using aqueous hydrosulphite in biphase, in alkaline medium in the presence of a water immiscible organic solvent. The hydroquinone of formula 4 was isolated by precipitating out the soild in presence of a hydrocarbon solvent. In the prior art process this reduction is effected in homogeneous phase using water miscible solvent. The modified process according to the present invention increases the yield of the reduced product of the formula 4 substantially (to about 96 % as compared to about 50% as per the prior art process). According to the improved process of the present invention , the brominated product compound of formula 14b was isolated by precipitating out the soild in presence of a hydrocarbon solvent. The process described above increases the yield of the brominated compound (to about 96 % as compared to 75 % as per the prior art process)
In the above mentioned process the purity of 2,3,4,5 tetramethoxy 6 methyl bromo benzene of the formula 14 b is enhanced when formed by first alkylation of 2,3 Dimethoxy 5 methyl 1,4 Hydroquinone of the formula 2 , to form 2,3,4,5 tetramethoxy toluene compound of formula 4b which can be purified easily by vacuum distillation.
In another embodiment of the present invention there is provided a process for the preparation of novel compound of the formula la9
25


Which comprises,
i) reacting Grignard reagent of formula 11a ,

with solanesyl bromide of the formula 3b9 in presence of cuprous halide in a solvent under

inert atmosphere at a temperature in the range of-5°C to25°C,
ii)Quenching the resulting reaction mixture obtained in step ( i) in ammonium chloride solution, and extracting the compound of the formula la 9 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula la 9 obtained in (ii) by column chromatography to obtain above 95% pure compound of formula la 9
This process has been shown in Scheme no 10 given above
26

In another embodiment of the present invention there is provided an improved process for the preparation of C0Q9 of the formula 1 9

Which comprises,
ii) reacting Grignard reagent of formula 11a,

with solanesyl bromide of the formula 3b9jn presence of cuprous halide in a solvent under

inert atmosphere at a temperature in the range of-5 C to 25 C,
ii)Quenching the resulting reaction mixture obtained in step ( i) in ammonium chloride solution, and extracting the novel compound of the formula la 9
27


in a solvent and evaporating the solvent,
iii) Purifying the crude novel compound of the formula la 9 obtained in (ii) by column chromatography to obtain above 95% pure compound of formula la 9
iv) Deprotecting the novel compound of formula la 9 by conventional method to obtain the Coenzyme Q 9 hydroquinone of the formula l_9H
v)Oxidising the Coenzyme Q 9 hydroquinone of the formula l_9H to obtain Coenzyme Q 9 of the formula l 9
vi))Crystallizing the Coenzyme Q 9 of the formula 1 9 obtained in (v) to obtain the yellow to orange Coenzyme Q 9 of the formula 19
This process has been shown in the combined reaction Scheme nos 10 & 10 a given above
According to yet another embodiment of the present invention , there is provided an improved process for the preparation of the novel compound of the formula lb 9

which comprises,
28

i) Reacting Grignard reagent compound of formula l1b.
with solanesyl bromide of the formula 3b9


in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the novel compound of formula lb 9 in a solvent and evaporating the solvent,
iii) Purifying the crude novel compound of the formula lb 9 obtained in ii) by column chromatography to obtain above 95% pure novel compound of formula lb 9
This process is shown in the reaction Scheme 11 given above
According to yet another embodiment of the present invention , there is provided an improved process for the preparation of compound of the formula CoQ 9 of the formula 19
29


which comprises,

i) Reacting Grignard reagent compound of formula l1b.

30
with solanesyl bromide of the formula 3b9



v)Crystallizing the compound of the formula l_9_purified by column chromatography using organic solvent(s) , to obtain the yellow to orange Coenzyme Q 9 of the formula 1 9
This process is shown in the combined reaction Schemes 11 & 11 a given above
In another embodimnent of the present invention there is provided an improved process for the prepartion of novel compound of the formula 19

Which comprises
i) Reacting Grignard reagent compound of formula 18

31
with solanesyl bromide of the formula 3b9


in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°Cto25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the compound of formula 19

in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 19 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 19
This process is shown in the Scheme 12
According to yet another embodiment of the present invention, there is provided an improved process for the preparation of CoQ 9 of the formula 1 9

Which comprises
i) Reacting Grignard reagent compound of formula 18
32


with solanesyl bromide of the formula 3b9


3b9
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°Cto25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the compound of formula 19

in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 19 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 19 iv) Deprotecting the compound of formula 19_ by conventional method to obtain novel compound of formula 20
33


v)Oxidising the compound of formula 20 to obtain Coenzyme Q 9 of the formula 1 9 vi))Crystallizing the Coenzyme Q 9 of the formula 1 9 obtained in (v) to obtain the yellow to orange Coenzyme Q 9 of the formula 1 9
This process is also shown in the combined reaction Schemes 12 & 12 a
In another embodiment of the present invention there is provided a n improved process for the preparation of compound of formula la 10 ,

la 10 which comprises,
34
i) Reacting Grignard reagent compound of formula 11a


with decaprenyl bromide compound of the formula 3b10


10

3b10
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of -50C to 25°C,
ii) Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula la 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula la 10 obtained in ii) by column chromatography to obtain above 95% pure compound of formula la 10
This reaction is shown in Scheme 13
In another embodiment of the present invention there is provided a n improved process for the preparation of COQ1oof the formula 1a 1o,













which comprises,
i) Reacting Grignard reagent compound of formula 11a






35

with decaprenyl bromide compound of the formula 3b10,




in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii) Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula la 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula la 10 obtained in ii) by column chromatography to obtain above 95% pure compound of formula la 10
iv) Deprotecting the compound of formula la 10 by conventional method to obtain Coenzyme Q 1o hydroquinone of the formula l 10 H

OH l l l 10 H
v)Oxidising the Coenzyme Q 1o hydroquinone of the formula l 10 H by known methods to obtain Coenzyme Q 1o of the formula 1 10
vi))Crystallizing the Coenzyme Q 1oof the formula 1 m obtained in (v) using organic solvent(s) to obtain the yellow to orange Coenzyme Q 1oof the formula 1 10
This process is also shown in the reaction Schemes 13 & 13 a
36

According to yet another embodiment of the present invention , there is provided an improved process for the preparation of compound of the formula lb 10
OMe

MeO
MeO
OMe

lb 10

which comprises,
i) Reacting Grignard reagent compound of formula l1b,


l1b
with decaprenyl bromide of the formula 3b10




3b10

in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula lb 10 in a solvent and evaporating the solvent,
37

iii) Purifying the crude compound of the formula lb 10 obtained in ii) by column chromatography to obtain above 95% pure compound of formula lb 10
This process is shown in the reaction Scheme 14 given above
According to yet another embodiment of the present invention , there is provided an improved process for the preparation of compound of the formula CoQ1oof the formula l 10



l 10which comprises,
i) Reacting Grignard reagent compound of formula l1b,
OMe
OMe
L1b
with decaprenyl bromide of the formula 3b10




3b10



in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
38

ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula lb 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula lb 10obtained in ii) by column
chromatography to obtain above 95% pure compound of formula lb 10
(iv) Oxidising the compound of formula lb 10 by conventional method to obtain
Coenzyme Q 10 compound of formula l 10
v)Purifying the compound of the formula l 10obtained in step (iv) by column
chromatography,
vi)Crystallizing the compound of the formula l 10purified by column
chromatography ising organic solvent(s) to obtain the yellow to orange Coenzyme Q
10 of the formula l 10
This process is shown in the combined reaction Schemes 14 & 14 a given above
In an embodiment of the present net invention there is provided an improved process for the preparation of novel compound of the formula 21_
OMe
OCH2OCH2OCH2OCH3

Which comprises

21

i) Reacting novel Grignard reagent compound of formula 18
OCH2OCH2CH2OCH3





39

with decaprenyl bromide of the formula 3b10




3b10

in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the novel compound of formula 21
MeO
MeO
OCH2OCH2CH2OCH3

OMe

21
n a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 21 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 21
This process is shown in the reaction Scheme 15
According to yet another embodiment of the present invention , there is provided an improved process for the preparation of CoQ10 of the formula l 1o
Which comprises
i) Reacting novel Grignard reagent compound of formula 18
40



with decaprenyl bromide of the formula 3b10


3b10
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the novel compound of formula 21
OCH2OCH2CH2OCH3

MeO
MeO
OMe

21
in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 21 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 21
iv) Deprotecting the compound of formula 21 by conventional method to obtain compound of the formula 22
41


OMe 22
v)Oxidising the compound of the formula 22 by known methods to obtain Coenzyme Q 10of the formula 1 1o
(vi) Crystallizing the Coenzyme Q 1o of the formula 1 1o obtained in (v) from organic solvent(s) to obtain the yellow to orange Coenzyme Q 1o the formula 1 1o
This process is also shown in the combined reaction Schemes 15 & 15 a
In a preferred embodiment of the present invention the various steps in the processes described above may be carried out as follows ,
Reduction of 2,3 Dimethoxy 5 methyl 1,4 benzoquinone, CoQo of the formula 2 , may be carried out by with sodium hydrosulphite in neutral or alkaline medium, preferably alkaline medium more preferably sodium hydroxide by dissolving CoQo in a water immiscible organic solvent like ether, aromatic hydrocarbons, chlorinated hydrocarbons more preferably chlorinated hydrocarbons like methylene chloride, ethylene chloride, preferably methylene chloride. Thus the reaction may be carried out in biphase, at a temperature in the range of 0° C to 30° C preferably, 10-20° C . Isolation of 2,3 Dimethoxy 5 methyl 1,4 Hydroquinone compound of the formula 4 , thus formed , may be carried out by acidifying the above reaction mixture, separating the organic phase and concentrating the organic phase. The concentrated organic phase may be added to aliphatic or aromatic hydrocarbon solvent like hexane, heptane, petroleum ether, preferably heptane to precipitate and filter the compound of formula 4
Bromination of 2,3 Dimethoxy-5- methyl 1,4 Hydroquinone compound of formula 4, may be carried out with bromine in the presence of a chlorinated hydrocarbon solvent
42

like methylene chloride and ethylenechloride at a tempertaure in the range of 0-30 C preferably at 10-20° C. Isolation of the brominated compound 2,3 Dimethoxy-5-methyl-6-bromol,4 hydroquinone compound of formula 13 thus formed, may be carried out by quenching the resulting reaction mixture in aqueous medium, separating and concentrating the organic phase. The concentrated liquid may be added to a hydrocarbon solvent preferably heptane to precipitate and filter 2,3 dimethoxy 5 methyl 6 bromo hydroquinone compound of formula 13.
Alkylation of 2,3 dimethoxy 5 methyl 6 bromo hydroquinone compound of the formula 13 may be carried out with methoxy ethoxy methyl chloride in the presence of metal hydride in aromatic hydrocarbons preferably toluene or an alkali metal alkoxide base like sodium methoxide, sodium ethoxide preferably sodium methoxide, in alcohol, at a temperature in the range of- 30°C to 30°C preferably 15-25°C. 2,3-dimethoxy-5-methyl-6-bromol,4 hydroquinone methoxyethoxymethyl ether compound of formula 14a thus formed, may be isolated by quenching the reaction mixture in alcohol or aqueous medium, extracting in solvent like ether, aromatic hydrocarbon, chlorinated hydrocarbons preferably methylene dichloride, and concentrating the solvent.
2,3 dimethoxy-5-methyl-6-bromo 1,4 hydroquinone bismethoxyethoxymathyl ether compound of formula 14a, 2,3,4,5-tetramethoxy-6-methyl-bromo benzene compound of formula 14b or 2,3,4 trimethoxy-5-bromo-6-methyl phenol compound of formula 16 may be converted to the Grignard reagent, as given in literature.
2,3, Dimethoxy 5 methyl 1,4 Hydroquinone compound of the formula 4_ may be alkylated using dimethylsulphate in acetone or in aqueous medium or in presence of alkali, preferably in aqueous medium in presence of alkali. The resulting product 2,3,4,5 tetramethoxy toluene compound of formula 4b , may be isolated by extracting in solvent and distilling out the solvent. The resultant residue may be distilled under vacuum at 0.2 - 10 mm Hg, preferably 0.5-0.8 mm Hg, to obtain the distilled 2,3,4,5 tetramethoxy toluene compound of formula 4b in more than 96% HPLC purity. 2,3,4,5 tetramethoxy toluene compound of formula 4b may be brominated as given above to form 2,3,4,5 tetramethoxy 6 methyl bromo benzene compound of formula 14b.
43

The coupling of the Grignard reagent of the formula 11a, or l1b or 18 with solanesyl bromide or decaprenyl bromide of the formula 3b9_or 3b10 may be carried out in the presence of cuprous halide like cuprous chloride, cuprous bromide or cuprous iodide preferably cuprous bromide. Grignard reagent may be used in equivalent amount or excess of the solanesyl bromide or decaprenyl bromide in molar ratio of 1:1 to 1:4 preferably 1: 1.1 to 1:2. The reaction may be carried out by adding the cuprous salt to the Grignard reagent and allowing to equilibrate for sufficient time. The copper salt is used in 1: 1 to 1:0.1 molar ratio of the Grignard reagent. The solanesyl bromide or decaprenyl bromide of the formula 3b9 or 3b10 dissolved in a solvent, may be added to the Grignard reagent at temperature range of -25 C to 25 C preferably at room temperature. The solvent used may be the same as used for the Grignard reagent or different like aromatic hydrocarbon aliphatic hydrocarbon like toluene, hexamethylphoshphoric triamide. The solvent for dissolving the solanesyl bromide or decaprenyl bromide may be preferably the same as used in Grignard reaction. The coupling of the Grignard reagent of the formula 11a, or l1b or 18 with solanesyl bromide or decaprenyl bromide of the formula 3b9_or 3b10 may also be carried out by adding cuprous salt to the solution of solanesyl bromide or decaprenyl bromide of the formula 3b9_or 3b10 and the Grignard reagent of the formula 11a, or l1b or 18 may be added to the above reaction mixture. The reaction may be monitored by HPLC and the rate of addition of the polyprenyl bromide solution may be adjusted with the rate of reaction. The reaction may be quenched in an aqueous medium in acidic or ammonium chloride solution preferably ammonium chloride solution, and the respective product of the formula la 9 or la 10, or la 9 or la 10 in or 19 or 21. may be extracted in an water immisicible solvent, solvent evaporated, and the crude compound may be purified by column chromatography to obtain more than 96 % pure compound..
Deprotection followed by oxidation of the coupled product of the formula la 9 or l a 10 or 19 or 21. may be carried out by method given in literature to obtain the final product of compound of formula 1_9or l 10.
44

The coupled product lb9 or lb 10 may be directly oxidized with eerie ammonium nitrate in acetonitrile as described in literature to obtain the final product of compound of formula l 9or lb 10
The details of the process are given in the Examples below which are provided for illustration only and therefore they should not be construed to limit the scope of the invention
Example 1
Preparation of Grignard reagent of 2,3 Dimethoxv-5-bromo-6-methyl 1,4 dimethoxyethoxy methyl ether compound of formula 11a.
2,3 dimethoxy 5-methyl 1,4 benzoquinone compound of formula 2, 2.5 g was dissolved in 7.5 ml of methylene dichloride and treated with sodium hydrosulphite(3.56 g) in an alkaline solution at 10-20 ° C. After 2 hours the reaction mixture was treated with cone. HC1 (3.4 ml) to acidic pH. The reaction mixture was extracted with methylene dichloride and washed with water. The organic solvent was concentrated and poured in hexane. The precipitated solid was filtered to obtain 2.25 g of 2,3 dimethoxy 5- methyl 1,4 hydroquinone compound of formula 4. The solid was taken in methylene dichloride and treated with bromine 1.96 g. at 10-20° C. The reaction was quenched in water after 2 hours and extracted in methylene dichloride . The methylene dichloride was evaporated. The concentrated mass was added to hexane to precipitate out the solid of 2,3 dimethoxy -5- bromo -6-methyl 1,4 hydrquinone 3.06 g. The bromo compound was dissolved in toluene and treated with 1.024 g sodium hydride (60% suspension) in toluene at 0 to -5 ° C. Methoxyethoxy methyl chloride 3.17 g was added at 5 to 10° C . The temperature was slowly raised to room temperature and the reaction was continued for 2 hrs. The reaction was quenched with methanol, followed by water and the toluene layer separated. The organic layer was distilled under vacuum to obtain 4.65 g of 2,3 Dimethoxy-5-bromo-6-methyl 1,4 hydroquinone di methoxyethoxy methyl ether compound of the formula 14a. The compound of formula 14a (4.65g) was reacted with Magnesium (0.30lg) in tetrahydrofuran, in presence of a pinch of iodine at ambient temperature to form the Grignard reagent of 2,3 dimethoxy-5-bromo-6-methyl 1,4 dimethoxyethoxy methyl ether compound of formula 11a
45

Example 2
Preparation of Grignard reagent of 2,3 Dimethoxy-5-bromo-6-methyl 1,4 dimethoxyethoxy methyl ether compound of formula 11a.
2,3 dimethoxy 5-methyl 1,4 benzoquinone compound of formula 2, 2.5 g was dissolved in 7.5 ml of methylene dichloride and treated with sodium hydrosulphite(3.56 g) in alkaline solution at 10 -20 ° C. After 2 hours the reaction mixture was treated with cone. HCl 3.4 ml to acidic pH. The reaction mixture was extracted with methylene dichloride and washed with water. The organic solvent was concentrated and poured in hexane 1oml. The precipitated solid was filtered to obtain 2.25 g of 2,3 dimethoxy 5 methyl 1,4 hydroquinone compound of formula 4. The solid was taken in methylene dichloride 15 ml and treated with bromine 1.96 g at 10 -20 ° C . The reaction was quenched in water after 2 hours and extracted in methylene dichloride . The methylene dichloride was evaporated. The concentrated mass was added to hexane to precipitate out the solid of 2,3 dimethoxy-5 bromo-6-methyl 1,4 hydrquinone 3.06 g. The bromo compound was dissolved in methanol and treated with sodium methoxide 1.5g at 5-10 C. Methoxyethoxy methyl chloride 3.17 g was added at 5 ° C - 10 C, the temperature raised to room temperature and maintained for 8 hrs. The reaction was quenched in water and extracted in diisopropyl ether. The organic layer was distilled under vacuum to obtain 4.75g of 2,3 Dimethoxy-5-bromo-6-methyl 1,4 di methoxyethoxy methyl ether compound of the formula 14a. The compound was reacted with Magnesium (0.34g) in tetrahydrofuran, in presence of a pinch of iodine at ambient temperature to form the Grignard reagent of 2,3 dimethoxy-5-bromo-6-methyl 1,4 dimethoxyethoxy methyl ether of the formula 11a.
Example 3
Preparation of Grignard reagent of 2, 3, 4, 5 tetramethoxy-6-methyl-bromobenzene compound of formula l1b
2,3dimethoxy-5-methyl 1,4 benzoquinone compound of formula 2j 2.5 g was dissolved in 7.5 ml of methylene dichloride and treated with sodium hydrosulphite (3.56 g) in alkaline solution at 10 -20 ° C. After 2 hours the reaction mixture was treated with cone. HCl (3.4 ml) to acidic pH. The reaction mixture was extracted with
46

methylene dichloride and washed with water. The organic solvent was concentrated and poured in hexane. The precipitated solid was filtered to obtain 2.25 g. of 2,3 dimethoxy 5 methyl 1,4 hydroquinone compound of formula 4. The solid was taken in alkaline solution and dimethyl sulphate 5.75 g was added at 40-50° C. The reaction mixture was quenched after 4 hours in water and extracted in methylene dichloride. The solvent was evaporated and the crude obtained was distilled under vacuum at 80 C at 0.5-1.0 mm Hg to obtain 2.33 g of 2,3, 4,5 tetramethoxy toluene. The compound was taken in methylene dichloride 15 ml and treated with bromine 1.75g. at 10 -20°C . The reaction was quenched in water after 2 hours and extracted in methylene dichloride. The methylene dichloride was evaporated. The concentrated mass was added to hexane to precipitate out the solid of 2,3,4,5 tetramethoxy-6-methyl bromobenzene, 3.03g, compound of formula 14b. The compound of formula 14b was reacted with Magnesium (0.30g) in tetrahydrofuran, at ambient temperature, in presence of a pinch of iodine to form the Grignard reagent of 2,3,4,5 tetramethoxy-6-methyl bromobenzene compound of the formula l1b.
Example 4
Preparation of Grignard reagent of 2, 3, 4, 5 tetramethoxy-6-methyl-bromobenzene compound of formula l1b
2,3 dimethoxy 5-methyl 1,4 benzoquinone compound of formula 2, 2.5 g was dissolved in 7.5 ml of methylene dichloride and treated with sodium hydrosulphite (3.56 g) in alkaline solution at 10 -20 ° C. After 2 hours the reaction mixture was treated with cone. HC1 (3.4 ml) to acidic pH. The reaction mixture was extracted with methylene dichloride and washed with water. The organic solvent was concentrated and poured in hexane. The precipitated solid was filtered to obtain 2.25 g. of 2,3 dimethoxy 5 methyl 1,4 hydroquinone compound of formula 4. The solid was taken in acetone, potassium carbonate (6.3 g) and dimethyl sulphate (5.75) g was added at 40-50° C. The reaction mixture was quenched after 4 hours in water and extracted in methylene dichloride. The solvent was evaporated and the crude obtained was distilled under vacuum at 80° C at 0.5-1.0 mm Hg to obtain 2.33 g of 2,3, 4,5 tetramethoxy toluene. The compound was taken in methylene dichloride 15 ml and treated with bromine 1.75 g. at 10 -20° C. The reaction was quenched in water after 2 hours and extracted in methylene dichloride . The methylene dichloride was
47

evaporated. The concentrated mass was added to hexane to precipitate out the solid of 2,3,4,5 tetramethoxy-6-methyl bromobenzene, 3.03g, compound of formula 14b. The compound 14b was reacted with Magnesium (0.30g) in tetrahydrofuran, at ambient temperature, in presence of a pinch of iodine to form the Grignard reagent of 2,3,4,5 tetramethoxy-6-methyl bromobenzene compound of the formula l1b.
Example - 5
Preparation of novel Grignard reagent of 2,3,4-trimethoxy-5-bromo-6-methyl-hydroquinone-1-methoxyethoxylmethyl ether of the formula 18.
2,3,4 trimethoxy-6- methyl-phenol compound of formula 15, 2.42g was taken in methylene dichloride 15 ml and treated with bromine 1.96 g at 10 -20 ° C. The reaction was quenched in water after 2 hours and extracted in methylene dichloride. The methylene chloride layer was evaporated. The concentrated mass was added to hexane to precipitate out the solid of 2,3,4 trimethoxy-5 bromo-6-methyl- phenol 3.22 g of formula 16. The bromo phenol of formula 16 was dissolved in toluene and treated with 0.513 g sodium hydride (60% suspension) in toluene at 0 to -5 ° C. Methoxyethoxy methyl chloride 1.59 g was added at 5 -10 ° C. The temperature was slowly raised to room temperature and maintained for 2 hrs. The reaction was quenched in water and the toluene layer separated. The organic layer was distilled under vacuum to obtain 4.03g of 2,3,4-trimethoxy-5-bromo-6-methyl-hydroquinone-1 -methoxyethoxylmethyl ether compound of the formula 17. The compound of formula 17 was reacted with Magnesium (0.35 g) in tetrahydrofuran, at ambient temperature, in presence of a pinch of iodine, to form the Grignard reagent of 2,3,4-trimethoxy-5-bromo-6-methyl-hydroquinone-l-methoxyethoxylmethyl ether of the formula 18.
1 H-NMR(300 MHz, CDC13, 2.33 (3H, -CH 3), 3.38-3.94 (18H, -OCH 20-, -OCH2 CH 2O-, -OCH 3)
Example 6
Preparation of novel Grignard reagent of 2,3,4-trimethoxy-5-bromo-6-methyl-hydroquinone-1-methoxyethoxylmethyl ether of the formula 18.
48

2,3,4 trimethoxy-6- methyl-phenol compound of formula 15, 2.42 g was taken in
methylene dichloride 15 ml and treated with bromine 1.96 g at 10 -20 ° C. The
reaction was quenched in water after 2 hours and extracted in methylene dichloride.
The methylene chloride layer was evaporated. The concentrated mass was added to
hexane to precipitate out the solid of 2,3,4 trimethoxy-5 bromo-6-methyl- phenol 3.22
g of formula 16. The bromo phenol of formula 16 was dissolved in in methanol and
treated with sodium methoxide 0.75 g at 5-10 C. Methoxyethoxy methyl chloride
1.59 g was added at5°C- 10°C and the temperature was raised to room temperature
and maintained for 8 hrs. The reaction was quenched in water and extracted in
diisopropyl ether. The solvent was distilled under vacuum to obtain 4.0 g of 2,3,4-
trimethoxy-5 -bromo-6-methyl-hydroquinone-1 -methoxyethoxylmethyl ether
compound of the formula 17. The compound of formula 17 was reacted with Magnesium (0.35 g) in tetrahydrofuran, at ambient temperature, in presence of a pinch of iodine, to form the Grignard reagent of 2,3,4-trimethoxy-5-bromo-6-methyl-hydroquinone-1-methoxyethoxylmethyl ether of the formula 18.
1 H-NMR(300 MHz, CDC13, 2.33 (3H, -CH 3), 3.38-3.94 (18H, -OCH 20-, -OCH2 CH 2O-, -OCH 3)
Example 7 Preparation of novel compound of the formula la 9
The Grignard reagent of 2,3 Dimethoxy-5-bromo-6-methyl 1,4 hydroquinone dimethoxyethoxy methyl ether of the formula 11a prepared by the process described in Example 1, was cooled to 0-5° C. Cuprous bromide (0.65g) was added to the Grignard solution of formula 11a, stirred at room temperature for 1 hour, followed by dropwise addition of a solution of solanesyl bromide in tetrahydrofuran (4 g in 25 ml tetrahydrofuran). The reaction mixture was stirred for four hours and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.2 g of crude, which was purified by column chromatography to give 4.4 g of the pure compound of formula la 9.
Example 8 Preparation of novel compound of formula la 9
49

The Grignard reagent of 2,3 Dimethoxy-5-bromo-6-methyl 1,4 dimethoxyethoxy methyl ether compound of the formula 11a prepared by the process described in Example 1, was slowly added to a solution of solanesyl bromide in tetrahydrofuran (4 g in 25 ml tetrahydrofuran) in presence of cuprous bromide (0.65 g). The reaction was continued for four hours at room temperature and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.8 g of crude, which was purified by column chromatography to give 4.0 g of the pure compound of formula la 9
Example 9 Preparation of novel compound of formula lb 9
The Grignard reagent of 2,3,4,5 tetramethoxy-6-methyl bromobenzene compound of the formula l1b, prepared by the process described in Example 3, was cooled to 0-5 C. Cuprous bromide (0.75g) was added to the Grignard solution of formula l1b, stirred at room temperature for 1 hour, followed by dropwise addition of a solution of solanesyl bromide in tetrahydrofuran (4g in 25 ml tetrahydrofuran). The reaction mixture was stirred for four hours and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.0 g of crude, which was purified by column chromatography to give 3.78 g of the pure compound of formula, lb 9.
Example 10 Preparation of novel compound of formula lb 9
The Grignard reagent of 2,3,4,5 tetramethoxy-6-methyl bromobenzene compound of the formula l1b, prepared by the process described in Example 3, was slowly added to a solution of solanesyl bromide in tetrahydrofuran (4 g in 25 ml tetrahydrofuran) in presence of cuprous bromide (0.75 g). The reaction was continued for four hours at room temperature and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and
50

evaporated to give 7.0g of crude, which was purified by column chromatography to give 3.36 g of the pure compound of formula lb 9
Example 11 Preparation of novel compound of formula 19
The Grignard reagent of 2,3,44rimethoxy-5-bromo-6-methyl-hydroquinone-l-methoxyethoxylmethyl ether of the formula 18 prepared by the process described in Example 5, was cooled to 0-5 C. Cuprous bromide (0.79g) was added to the Grignard solution of formula 18, stirred at room temperature for 1 hour, followed by dropwise addition of a solution of solanesyl bromide in tetrahydrofuran (4 g in 25 ml tetrahydrofuran). The reaction mixture was stirred for four hours and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.2 g of crude, which was purified by column chromatography to give 4 g of the pure compound of formula 19.
Example 12 Preparation of novel compound of formula 19
The Grignard reagent of 2, 3, 4 - trimethoxy - 5 - bromo - 6 - methyl - hydroquinone-1-methoxyethoxylmethyl ether of the formula 18 prepared by the process described in Example 5, was slowly added to a solution of solanesyl bromide in tetrahydrofuran (4g in 25 ml tetrahydrofuran) in presence of cuprous bromide (0.79 g). The reaction was continued for four hours at room temperature and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.8 g of crude, which was purified by column chromatography to give 3.68g of the pure compound of formula
19
Example 13 Preparation of compound of the formula la 10
51

The Grignard reagent of 2,3 Dimethoxy-5-bromo-6~methyl 1,4 hydroquinone dimethoxyethoxy methyl ether of the formula 11a prepared by the process described in Example 1, was cooled to 0-5 C. Cuprous bromide (0.65g) was added to the Grignard solution of formula 11a, stirred at room temperature for 1 hour, followed by dropwise addition of a solution of decaprenyl bromide in tetrahydrofuran (4.39 g in 25 ml tetrahydrofuran). The reaction mixture was stirred for four hours and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.2 g of crude, which was purified by column chromatography to give 4.39 g of the pure compound of formula la 10.
Example 14 Preparation of compound of formula la 10
The Grignard reagent of 2,3 Dimethoxy-5bromo-6-methyl 1,4 dimethoxyethoxy methyl ether compound of the formula 11a prepared by the process described in Example 1, was slowly added to a solution of decaprenyl bromide in tetrahydrofuran (4.39g in 25ml tetrahydrofuran) in presence of cuprous bromide (0.65 g). The reaction was continued for four hours at room temperature and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.8 g of crude, which was purified by column chromatography to give 3.88 g of the pure compound of formula la 10
Example 15 Preparation of compound of formula lb in
The Grignard reagent of 2,3,4,5 tetramethoxy-6-methyl bromobenzene compound of the formula l1b, prepared by the process described in Example 3, was cooled to 0-5° C. Cuprous bromide (0.75g) was added to the Grignard solution of formula l1b, stirred at room temperature for 1 hour, followed by dropwise addition of a solution of decaprenyl bromide in tetrahydrofuran (4.39g in 25 ml tetrahydrofuran). The reaction mixture was stirred for four hours and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous
52

sodium sulphate and evaporated to give 7.0 g of crude, which was purified by column chromatography to give 4.11 g of the pure compound of formula, lb 10.
Example 16 Preparation of compound of formula lb 10
The Grignard reagent of 2,3,4,5 tetramethoxy-6-methyl bromobenzene compound of the formula l1b, prepared by the process described in Example 3, was slowly added to a solution of decaprenyl bromide in tetrahydrofuran (4.39 g in 25 ml tetrahydrofuran) in presence of cuprous bromide (0.75 g). The reaction was continued for four hours at room temperature and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.0g of crude, which was purified by column chromatography to give 3.65 g of the pure compound of formula lb 10
Example 17 Preparation of novel compound of formula 21
The Grignard reagent of 2, 3, 4 - trimethoxy - 5 - bromo - 6 - methyl - hydroquinone-1 -methoxyethoxylmethyl ether of the formula 18 prepared by the process described in Example 5, was cooled to 0-5° C. Cuprous bromide (0.79g) was added to the Grignard solution of formula 18, stirred at room temperature for 1 hour, followed by dropwise addition of a solution of decaprenyl bromide in tetrahydrofuran (4.39 g in 25 ml tetrahydrofuran). The reaction mixture was stirred for four hours and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.2 g of crude, which was purified by column chromatography to give 4.45 g of the pure compound of formula 21.
Example 18 Preparation of novel compound of formula 21
The Grignard reagent of 2, 3, 4 - trimethoxy - 5 - bromo - 6 - methyl - hydroquinone-1-methoxyethoxylmethyl ether of the formula 18 prepared by the process described
53

in Example 5, was slowly added to a solution of decaprenyl bromide in tetrahydrofuran (4.39g in 25 ml tetrahydrofuran) in presence of cuprous bromide (0.79 g). The reaction was continued for four hours at room temperature and the mixture quenched in 5% ammonium chloride solution and extracted in diethyl ether. The solvent was dried over anhydrous sodium sulphate and evaporated to give 7.8 g of crude, which was purified by column chromatography to give 3.95 g of the pure compound of formula 21
Example 19 Preparation of compound of formula l9 from novel compound la9
The compound of the formula la g (4.4 g )prepared by the process described_in Example 7 was treated with 48% HBr solution (0.22 ml), in presence of isopropanol for 4 hours. The isopropanol was distilled off and the residue was taken in n - hexane . The hexane solution was washed with water dried over anhydrous sodium sulphate and distilled under vacuum to obtain 3.56 g of the residue, C0Q9 dihydroquinone compound of formula 19 H. The compound of formula 19 H was oxidized with ferric chloride 2.56 g in 1ml water, in presence of isopropanol at room temperature for 3 hours. The reaction was quenched in water and extracted in hexane. The hexane layer was dried over anhydrous sodium sulphate and evaporated to give crude CoQ 9. The crude CoQ 9 was crystallized in ethanol, at 10 - 15° C, to obtain 2.67 g of pure compound, with overall yield from solanesyl bromide as 58%.
Example 20
Preparation of CoQ9 of formula 19 from novel compound lb9
The compound of the formula lb 9_(3.78 g) prepared by the process described in Example 9 was taken in 48 ml of methylene dichloride and treated with a solution 4 g of eerie ammonium nitrate in 25 ml of acetonitrile and 25 ml of water at 0° C. The reaction mixture was quenched in water and extracted in methylene dichloride solution. The methylene dichloride was concentrated under vacuum to obtain crude CoQ 9. The crude CoQ 9 was purified by column chromatography and crystallized in ethanol, at 10-15 C to obtain 2.34 g of pure compound, with overall yield from solanesyl bromide as 51 %.
54

Example 21
Preparation of compound of formula U from novel compound 19 through novel compound 20
The compound of the formula 19 (4.0 g )prepared by the process described in Example 11 was treated with 48% HBr solution (0.22 ml), in presence of isopropanol for 4 hours. The isopropanol was distilled off and the residue was taken in n - hexane . The hexane solution was washed with water dried over anhydrous sodium sulphate and distilled under vacuum to obtain 3.24 g of the residue, novel compound of formula 20. The compound of formula 20 was oxidized with ferric chloride 2.56 g in l ml water, in presence of isopropanol at room temperature for 3 hours. The reaction was quenched in water and extracted in hexane. The hexane layer was dried over anhydrous sodium sulphate and evaporated to give crude CoQ 9. The crude CoQ 9 was crystallized in ethanol, at 10 - 15 C, to obtain 2.30 g of pure compound, with overall yield from solanesyl bromide as 50 %.
Example 22 Preparation of compound of formula l 10 From la 10
The compound of the formula la 10 (4.39 g )prepared by the process described in Example 13 was treated with 48% HBr solution (0.22 ml), in presence of isopropanol for 4 hours. The isopropanol was distilled off and the residue was taken in n - hexane . The hexane solution was washed with water dried over anhydrous sodium sulphate and distilled under vacuum to obtain 3.56 g of the residue, C0Q10 dihydroquinone compound of formula 1 10 H. The compound of formula 1 10 H was oxidized with ferric chloride 2.56 g in l ml water, in presence of isopropanol at room temperature for 3 hours. The reaction was quenched in water and extracted in hexane. The hexane layer was dried over anhydrous sodium sulphate and evaporated to give crude CoQ 10. The crude CoQ 10 was crystallized in ethanol, at 10 - 15° C, to obtain 2.53 g of pure compound, with overall yield from decaprenyl bromide as 51 %.
Example 23 Preparation of CoQ 10 of formula 1 10 in from l b 10
55

The compound of the formula lb 10 (4.11 g) prepared by the process described in Example 15 was taken in 48 ml of methylene dichloride and treated with a solution 4 g of eerie ammonium nitrate in 25 ml of acetonitrile and 25 ml of water at 0° C. The reaction mixture was quenched in water and extracted in methylene dichloride solution. The methylene dichloride was concentrated under vacuum to obtain crude CoQ 1o. The crude CoQ 1o was purified by column chromatography and crystallized in ethanol, at 10-15° C, to obtain 2.54 g of pure compound, with overall yield from decaprenyl bromide as 51.0 %.
Example 24 Preparation of CoQ 10 of formula l 10 from 21
The compound of the formula 21 (4.45 g )prepared by the process described in Example 17 was treated with 48% HBr solution (0.22 ml), in presence of isopropanol for 4 hours. The isopropanol was distilled off and the residue was taken in n - hexane . The hexane solution was washed with water dried over anhydrous sodium sulphate and distilled under vacuum to obtain 3.89 g of the residue, compound of formula 22. The compound of formula 22 was oxidized with ferric chloride 2.56 g in 1ml water, in presence of isopropanol at room temperature for 3 hours. The reaction was quenched in water and extracted in hexane. The hexane layer was dried over anhydrous sodium sulphate and evaporated to give crude CoQ io. The crude CoQ1o was crystallized in ethanol, at 10 - 15° C, to obtain 2.77 g of pure compound, with overall yield from decaprenyl bromide as 55.8 %.
Advantages of the invention
1. Provides Straight forward coupling of the "benzoquinone nucleus" with the "poly prenyl side chain" for the preparation of the coenzymes Q namely, CoQ 9 and CoQ 10
2 Provides stereoselective coupling reaction for preparation of coenzymes Q namely, CoQ 9 and CoQ 10 by simple Grignard reaction, maintaining the geometrical isomer of the double bond. Controlling cis isomer in the reaction decreases
56

purification loss incurred in removing unwanted cis isomer, thereby making the process cost effective.
if,
3. Provides a novel Grignard reagent compound of formula 18 and its preparation , which is useful for the preparation of Coenzymes namely, CoQ 9 and CoQ 10.
4. Provides novel intermediates compounds of formulae la 9, lb 9, 19 and 20 useful for the preparation of CoQ 9
5. Provides novel intermediate compounds of formula 21 useful for the preparation of CoQ 10
57

We Claim
1. A novel Grignard reagent of the formula 18



2. A process for the preparation of novel Grignard reagent of the Formula 18






which comprises,
(i) Brominating the compound of the formula 15


MeO
MeO

by known method, to obtain compound of formula 16

16

(ii) Alkylating the compound of the formula 16 obtained in step (ii ) with methoxyethoxymethyl chloride in the presence of a base, an alkali metal alkoxide or metal hydride,to obtain2,3-dimethoxy-5-methyl-6-bromo hydroquinonel,4 dimethoxyethoxymethyl ether compound of formula 17
OCH2OCH2CH2OCH3

17
(iii)Reacting the compound of the formula 17 obtained in step (ii) with Magnesium
in presence of ether at 0-65 ° C, in presence of iodine and dibromoethane, to obtain
the novel Grignard reagent of the formula 18
(iv) cooling the resulting reaction mixture to room temperature , filtering to get the
novel Grignard reagent of the formula 18 in solution .
3. An improved process for the preparation of Grignard reagent of the formula
11a,



which comprises






i Reducing 2,3 dimethoxy -5 -methyl 1,4 benzoquinone (CoQo ) of the





59

with sodium hydrosulphite, in alkaline medium, in biphase, in the presence of a water
immiscible organic solvent, and
ii separating the organic phase,
iii evaporating the oraganic phase obtained in step ii to obtain a concentrated



residue compound of formula 4
iv Adding the concentrated residue obtained in step iii to a hydrocarbon solvent to
precipitateout the soild of the compound of formula 4and filtering out the solid
v Brominating the resulting compound of the formula 4 with bromine in
chlorinated hydrocarbon at 0-25 C ,
vi Quenching the resultant reaction mixture in step (i) in aqueous medium to
obtain aqueous and organic phase and separating the oraganic phase,
vii Evaporating the organic phase obtained in step ii to obtain a concentrated
residue of 2,3 dimethoxy -5 -methyl -6-bromo 1,4 hydroquinone of the formula 13





viii Adding the concentrated residue obtained in step vii to a hydrocarbon solvent
to precipitate out the soild of the compound of formula 13 and filtering out the solid
ix Alkylating the 2,3 dimethoxy -5 -methyl -6-bromo 1,4 hydroquinone of the
formula 13 obtained in step (viii) with methoxyethoxymethyl chloride in the


presence of a base, an alkali metal alkoxide or metal hydride, to obtain 2,3-dimethoxy-5-methyl-6-bromo hydroquinonel,4 dimethoxyethoxymethyl ether compound of formula 14a,








and x Reacting the compound of the formula 14a obtained in step (ix ) with Magnesium
in presence of ether, iodine and dibromoethane, at a temperature in the range of 0-65
C, to obtain the Grignard reagent of the formula 11 a
4. An improved process for the preparation of Grignard reagent of the Formula l1b.



which comprises,
i. Reducing 2,3 dimethoxy-5-methyl 1,4 benzoquinone (CoQo) of the formula 2





With sodium hydrosulphite, in alkaline medium, in biphase, in the presence of a water
immiscible organic solvent, and
ii separating the organic phase,
iii evaporating the oraganic phase obtained in step ii to obtain a concentrated
residue of compound of formula 4






iv Adding the concentrated residue obtained in step iii to a hydrocarbon solvent to precipitate out the soild of the compound of formula 4and filtering out the solid v.Alkylating the compound of the formula 4_, with alkyl sulphate by known method to obtain 2,3,4,5 tetramethoxy toluene compound of formula 4b,









viBrominating the resulting compound of the formula 4b with bromine in
chlorinated hydrocarbon at 0-25 C ,
vii Quenching the resultant reaction mixture in step (vi) in aqueous medium to obtain
aqueous and organic phase and separating the oraganic phase,
viiiEvaporating the organic phase obtained in step vii to obtain a concentrated residue
of 2,3,4,5 tetramethoxy 6-bromo toluene of the formula 14b
OMe

ix Adding the concentrated residue obtained in step viii to a hydrocarbon solvent to precipitate out the soild of the compound of formula 14b and filtering out the solid


x Reacting the compound of the formula 14b obtained in step (iii) with Magnesium in presence of ether, iodine and dibromoethane at 0-65 C, to obtain the Grignard reagent of the formula l1b










5. A novel compound of the formula la 9
0CH20CH2CH20CH3

MeO
MeO
0CH20CH2CH20CH3

la 9
useful for the preparation of coenzyme, namely, CoQ 9
6. A process for the preparation of novel compound of the formula la 9






Which comprises,
iii) reacting Grignard reagent of formula 11a
63




with solanesyl bromide of the formula 3b9 presence of cuprous halide in a solvent under

3b 9
inert atmosphere at a temperature in the range of-5 C to 25 C,
ii)Quenching the resulting reaction mixture obtained in step ( i) in ammonium chloride solution, and extracting the compound of the formula la 9 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula la 9 obtained in (ii) by column chromatography to obtain above 95% pure compound of formula la 97. An improved process for the preparation of CoQ 9 of the formula 19

64
Which comprises,

i) reacting Grignard reagent of formula 11a



with solanesyl bromide of the formula 3b9 in presence of cuprous halide in a solvent under



3b 9
inert atmosphere at a temperature in the range of-5° C to 25° C,
ii)Quenching the resulting reaction mixture obtained in step ( i) in ammonium chloride solution, and extracting the novel compound of the formula la 9g


MeO
MeO

la 9
in a solvent and evaporating the solvent,
iii) Purifying the crude novel compound of the formula la 9 obtained in (ii) by column chromatography to obtain above 95% pure compound of formula la 9
iv) Deprotecting the novel compound of formula la 9 by conventional method to obtain the Coenzyme Q 9 hydroquinone of the formula l 9 H
65

v)Oxidising the Coenzyme Q 9 hydroquinone of the formula 1 9 H to obtain Coenzyme Q 9 of the formula 19
vi))Crystallizing the Coenzyme Q 9 of the formula 1 9 obtained in (v) to obtain the yellow to orange Coenzyme Q 9 of the formula 19
8. A novel compound of formula novel of the formula lb 9
OMe

MeO
MeO
OMe

l b 9
9. A process for the preparation of novel compound of the formula lb 9
OMe

MeO
MeO
OMe

l b_9
which comprises,
i) Reacting Grignard reagent compound of formula l1 b.





with solanesyl bromide of the formula 3b9

66




in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the novel compound of formula lb 9 in a solvent and evaporating the
solvent,
iii) Purifying the crude novel compound of the formula lb 9 obtained in ii) by
column chromatography to obtain above 95% pure novel compound of formula lb 9
10. An improved process for the preparation of compound of the formula CoQ 9 of the formula 1_9

which comprises,
i) Reacting Grignard reagent compound of formula lib,




with solanesyl bromide of the formula 3b9
67




in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the novel compound of formula lb 9 in a solvent and evaporating the
solvent,
iii) Purifying the crude novel compound of the formula lb 9 obtained in ii) by
column chromatography to obtain above 95% pure novel compound of formula lb 9
(iv) Oxidising the compound of formula lb 9 by conventional method to obtain
Coenzyme Q 9 compound of formula l 9
v)Purifying the compound of the formula l 9 obtained in step (v) by column
chromatography,
vi)Crystallizing the compound of the formula l 9 purified by column chromatography
using organic solvent(s) , to obtain the yellow to orange Coenzyme Q 9 of the
formula 19
11 A novel compound of the formula 19


12 A process for the preparation of the novel compound of formula 19
68

OCH2OCH2OCH2OCH3

MeO
MeO-^^^^
OMe

19
Which comprises
i) Reacting Grignard reagent compound of formula 18
OCH2OCH2CH2OCH3

OMe
18
with solanesyl bromide of the formula 3b9

3b9
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the compound of formula 19
69

OCH2OCH2CH2OCH3

MeO
MeO
OMe
19

in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 19 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 19
13 An improved process for the preparation of CoQ 9 of the formula 19






which comprises
i) Reacting Grignard reagent compound of formula 18






18
with solanesyl bromide of the formula 3b9
70





in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the compound of formula 19







in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 19 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 19 iv) Deprotecting the compound of formula 19_ by conventional method to obtain novel compound of formula 20

OMe
20
v)Oxidising the compound of formula 20 to obtain Coenzyme Q 9 of the formula 19

vi))Crystallizing the Coenzyme Q 9 of the formula 1 9 obtained in (v) to obtain the yellow to orange Coenzyme Q 9 of the formula 19
14 An improved process for the preparation of compound of formula la 10 ,



i) Reacting Grignard reagent compound of formula 11a






with decaprenyl bromide compound of the formula 3b10

Br 3b10
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii) Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula l a 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula l a 10 obtained in ii) by column chromatography to obtain above 95% pure compound of formula l a 10
15 An improved process for the preparation of COQ 10 of the formula 1 10,
72









which comprises,
ii) Reacting Grignard reagent compound of formula 11a



MeO
MeO
OCH2OCH2CH2OCH3

11a
with decaprenyl bromide compound of the formula


10


in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii) Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula la 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula la 10 obtained in ii) by column chromatography to obtain above 95% pure compound of la 10
73

iv) Deprotecting the compound of formula la 10 by conventional method to obtain Coenzyme Q 10 hydroquinone of the formula l 10 H


MeO
MeO

l 10 H
v)Oxidising the Coenzyme Q 10 hydroquinone of the formula l 10 H by known methods to obtain Coenzyme Q1oof the formula 1 10
vi))Crystallizing the Coenzyme Q1oof the formula 1 10 obtained in (v) using organic solvent(s) to obtain the yellow to orange Coenzyme Q1oof the formula 1 1o
16 An improved process for the preparation of compound of the formula lb 10
OMe

MeO
MeO
OMe

l b 10

which comprises,
i) Reacting Grignard reagent compound of formula l1b,



74

with decaprenyl bromide of the formula 3b10



3b10
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula lb 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula lb 10 in obtained in ii) by column chromatography to obtain above 95% pure compound of formula lb 10
17 An improved process for the preparation of Coenzyme Q 10 of the formula 1
10
O

l 10
which comprises,
i) Reacting Grignard reagent compound of formula l1 b,



with decaprenyl bromide of the formula 3b10
75







in presence of cuprous halide in a solvent under inert atmosphere at a temperature in the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and extracting the compound of formula lb 10 in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula lb 10obtained in ii) by column chromatography to obtain above 95% pure compound of formula lb 10
(iv) Oxidising the compound of formula lb 10by conventional method to obtain Coenzyme Q 1o compound of formula l 1o
v)Purifying the compound of the formula l 1o
obtained in step (iv) by column chromatography,
vi)Crystallizing the compound of the formula l 1o
purified by column chromatography ising organic solvent(s) to obtain the yellow to orange Coenzyme Q 1o of the formula 1 10
18. A novel compound of the formula 21
OCH2OCH2OCH2OCH3

OMe
21
19 A process for the preparation of compound of the formula 21.
76

OCH2OCH2OCH2OCH3

OMe

Which comprises

21

i) Reacting novel Grignard reagent compound of formula 18



with decaprenyl bromide of the formula 3b10



in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the novel compound of formula 21
MeO

MeO
OCH2OCH2CH2OCH3

OMe
21
77

n a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 21 obtained in ii) by column chromatography to obtain above 95% pure compound of formula 21
20 An improved process for the preparation of CoQio of the formula 1 10

l 10
which comprises
i) Reacting novel Grignard reagent compound of formula 18




with decaprenyl bromide of the formula 3b10

3b10
in presence of cuprous halide in a solvent under inert atmosphere at a temperature in
the range of-5°C to 25°C,
ii)Quenching the resulting reaction mixture of i) in ammonium chloride solution, and
extracting the novel compound of formula 21.
78


OCH2OCH2CH2OCH3
21
in a solvent and evaporating the solvent,
iii) Purifying the crude compound of the formula 21_ obtained in ii) by column chromatography to obtain above 95% pure compound of formula 21
iv) Deprotecting the compound of formula 21 by conventional method to obtain compound of the formula 22

v)Oxidising the compound of the formula 22 by known methods to obtain Coenzyme Q 10of the formula 1 10
(vi) Crystallizing the Coenzyme Q 10 of the formula 1 10obtained in (v) from organic solvent(s) to obtain the yellow to orange Coenzyme Q 10of the formula 1 10
21. An improved process as claimed in claims 3 & 4 wherein the reduction of 2,3 Dimethoxy 5 methyl 1,4 benzoquinone, CoQo of the formula 2 , is carried out using sodium hydrosulphite in neutral or alkaline medium, preferably alkaline medium more preferably sodium hydroxide at a temperature in the range of 0° C to 20° C preferably, 10-20°C.
79

22 An improved process as claimed in claims 3, 4 &21 wherein the reduction is carried out in a water immiscible organic solvent like ether, aromatic hydrocarbons, chlorinated hydrocarbons more preferably chlorinated hydrocarbons like methylene chloride, ethylene chloride, preferably methylene chloride.
23. A process as claimed in claims 3,4, 21 &22 wherein the isolation of 2,3 Dimethoxy 5 methyl 1,4 Hydroquinone compound of the formula 4_ is effected by acidifying the above reaction mixture, separating the organic phase and concentrating the organic phase.and adding the concentrated residue to aliphatic or aromatic hydrocarbon solvent like hexane, heptane, petroleum ether, preferably heptane to precipitate and filter the compound of formula 4
24 An improved process as claimed in claims 3,4 wherein the bromination of 2,3 Dimethoxy 5 methyl 1,4 Hydroquinone compound of formula 4 is carried out using bromine in the presence of a chlorinated hydrocarbon solvent like methylene chloride and ethylenechloride at a tempertaure in the range of 0-30 C preferably at 10-20 C
25 An improved process as claimed in claims 3,4 & 24 wherein the isolation of the brominated compound 2,3 Dimethoxy-5-methyl-6-bromol,4 hydroquinone compound of formula 13 formed is carried out by quenching the resulting reaction mixture in aqueous medium, separating and concentrating the organic phase at a temperature in the range of 0 to 20 ° C preferably at 0-5 ° C and adding the concentrated residue to aliphatic or aromatic hydrocarbon solvent like hexane, heptane, petroleum ether, preferably heptane to precipitate and filter the compound of formula 13
26. An improved process as claimed in claim 3 wherein the alkylation of 2,3 dimethoxy 5 methyl 6 bromo hydroquinone compound of the formula 13 is carried out using methoxy ethoxymethyl chloride in the presence of metal hydride in aromatic hydrocarbons preferably toluene or an alkali metal alkoxide base like sodium methoxide , sodium ethoxide preferably sodium methoxide, in alcohol, at a temperature in the range of - 30 °C to 30°C preferably 15-25°C.
80

27. An improved process as claimed in claims 3 & 26 wherein the 2,3 dimethoxy-5-methyl 6-bromo 1,4 hydroquinone methoxyethoxymathyl ether compound of formula 14a formed is isolated by quenching the reaction mixture in aqueous medium, extracting in solvent like ether, aromatic hydrocarbon, chlorinated hydrocarbons preferably methylene dichloride, and concentrating the solvent.
28 An improved process as claimed in claims 3,4 wherein the 2,3 dimethoxy 5 methyl 6 bromo 1,4 hydroquinone methoxyethoxymathyl ether compound of formula 14a, or 14b ,or 16 I is converted to the respective Grignard reagent, by reacting with Magnesium by conventional method
29 An improved process as claimed in claims 4 wherein Dimethoxy 5 methyl 1,4 Hydroquinone compound of the formula 4_ is alkylated using dimethylsulphate in acetone or in aqueous medium in presence of alkali preferably in aqueous medium in presence of alkali.
30. A process as claimed in claims 4 & 29 wherein the resulting 2,3,4,5 tetramethoxy toluene compound of formula 4b is isolated by extracting in solvent and distilling out the solvent, and the resulting residue is distilled under vacuum at 0.2 - lomm Hg, preferably 0.5-0.8 mm Hg,
31 An improved process as claimed in claim 6 to 7 & 9 to 18 & 20 wherein the coupling of the Grignard reagent of the formula 11a, or l1b or 18 with solanesyl bromide or decaprenyl bromide of the formula 3b9 or 3b10 is carried out in the presence of cuprous halide like cuprous chloride, cuprous bromide or cuprous iodide preferably cuprous bromide.
32 An improved process as claimed in claims 6 to 7 &9 to 18 & 20 & 31 wherein the Grignard reagent used is in excess of the solanesyl bromide or decaprenyl bromide , in a molar ratio of 1:1 to 1:4 preferably 1:1.1 to 1:2
33. An improved process as claimed in claim 6 to 7 & 9 to 18 & 20 & 31 & 32 wherein the copper salt is used in 1: 1 to 1:0.1 molar ratio of the Grignard reagent.
81

34 An improved process as claimed in claim 6 to 7 & 9 to 18 & 20 & 31 to 33 wherein the coupling of the Grignard reagent of 11a or l1b or 18 with solanesyl bromide or decaprenyl bromide is carried out at temperature range of -25 to 25 C preferably at room temperature
35 An improved process as claimed in claim 6 to 7 & 9 to 18 & 20 & 31 & 32 wherein the coupled product of the formula la 9 or la 10>r lb 9 or lb 10 or 19 or 21 is extracted in an water immmisicible solvent and solvent evaporated and crude purified by column chromatography

36. An improved process as claimed in claim 10 & 17 wherein the coupled product of the formulae lb9 or lb 10 is directly oxidized with eerie ammonium nitrate in acetonitrile by known methods to obtain the compound of formulae 1 9 or 1 10
37. An improved process as claimed in claim 7,13, 15,19 wherein the coupled product of the formulae la9 or la 10 or 19 or 21 is deprotected and oxidized by known methods to obtain the compound of formulae 1 9 or 1 10.

38 Novel Grignard reagent of the formula 18 substantially as described with particular reference to the Example 5 and 6
39 Novel compound of the formula la 9 substantially as described with particular reference to the Example 7 and 8
40 Novel compound of the formula lb 9 substantially as described with particular reference to the Example 9 and 10
41 Novel compounbd of the formula 19 substantially as described with particular reference to the Example 1 land 12
42 Novel compounbd of the formula 21. substantially as described with particular reference to the Example 17 and 18
82

43 An improved process for the preparaion of compounds of the formulae 11a & l1b substantially as described with particular reference to the Example 1 to 4
44 An improved process for the preparaion of compounds of the formulae la1o & lb10 substantially as described with particular reference to the Examples 13 to 16
45 An improved process for the preparation of CoQ 9 and C0Q10 substantially as described with particular reference to the Examples 19 to 24

83

Abstract

The present invention relates to novel intermediates for the preparation of coenzymes, processes for the preparation of the intermediates and an improved process for the preparation of Coenzymes. The present invention particularly relates to an improved process for the preparation of Coenzyme Q, more particularly for Conenzyme Q 9 and Coenzyme Q 10 for the preparation of Coenzyme Q9 and Coenzyme Q 10 of the formula l n.

Documents:

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805-MUM-2005-FORM 1(14-7-2005).pdf

805-MUM-2005-FORM 1(6-7-2005).pdf

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abstract1.jpg


Patent Number 247945
Indian Patent Application Number 805/MUM/2005
PG Journal Number 23/2011
Publication Date 10-Jun-2011
Grant Date 06-Jun-2011
Date of Filing 06-Jul-2005
Name of Patentee PIRAMAL HEALTHCARE LIMITED
Applicant Address PIRAMAL TOWER, GANPATRAO KADAM MARG, LOWER PAREL, MUMBAI-400013
Inventors:
# Inventor's Name Inventor's Address
1 ABHAY UPARE NICHOLAS PIRAMAL INDIA LTD, NICHOLAS PIRAMAL RESEARCH CENTRE, 1A, 1B & 1C - NIRLON COMPLEX, OFF WESTERN EXPRESS HIGHWAY, NEAR NSE COMPLEX, GOREGAON (E), MUMBAI - 400 063.
2 NITIN YASHWANT PAWAR NICHOLAS PIRAMAL INDIA LTD, NICHOLAS PIRAMAL RESEARCH CENTRE, 1A, 1B & 1C - NIRLON COMPLEX, OFF WESTERN EXPRESS HIGHWAY, NEAR NSE COMPLEX, GOREGAON (E), MUMBAI - 400 063.
3 GANESH WAGH NICHOLAS PIRAMAL INDIA LTD, NICHOLAS PIRAMAL RESEARCH CENTRE, 1A, 1B & 1C - NIRLON COMPLEX, OFF WESTERN EXPRESS HIGHWAY, NEAR NSE COMPLEX, GOREGAON (E), MUMBAI - 400 063.
4 MITA ROY NICHOLAS PIRAMAL INDIA LTD, NICHOLAS PIRAMAL RESEARCH CENTRE, 1A, 1B & 1C - NIRLON COMPLEX, OFF WESTERN EXPRESS HIGHWAY, NEAR NSE COMPLEX, GOREGAON (E), MUMBAI - 400 063.
5 AMIT CHAVAN NICHOLAS PIRAMAL INDIA LTD, NICHOLAS PIRAMAL RESEARCH CENTRE, 1A, 1B & 1C - NIRLON COMPLEX, OFF WESTERN EXPRESS HIGHWAY, NEAR NSE COMPLEX, GOREGAON (E), MUMBAI - 400 063.
6 HARIHARAN SIVARAMAKRISHNAN NICHOLAS PIRAMAL INDIA LTD, NICHOLAS PIRAMAL RESEARCH CENTRE, 1A, 1B & 1C - NIRLON COMPLEX, OFF WESTERN EXPRESS HIGHWAY, NEAR NSE COMPLEX, GOREGAON (E), MUMBAI - 400 063.
PCT International Classification Number A61K31/00
PCT International Application Number N/A
PCT International Filing date
PCT Conventions:
# PCT Application Number Date of Convention Priority Country
1 NA