Process for producing N-formylleucine of high purity

Organic compounds -- part of the class 532-570 series – Organic compounds – Carboxylic acids and salts thereof

Reexamination Certificate

Rate now

  [ 0.00 ] – not rated yet Voters 0   Comments 0

Details

C562S553000

Reexamination Certificate

active

06294692

ABSTRACT:

The invention relates to a process for producing N-formylleucine of high purity.
N-Formylleucine, composed of the leucine molecule with one of the hydrogen atoms attached to the nitrogen replaced by a formyl group, is a known compound used in particular in carrying out peptide syntheses.
Various processes for the formylation of amines, in particular of the amine groups of amino acids, are known. Industrially, amino acids are generally N-formylated by the action of formic acid or of an alkali metal formate. However, these processes have disadvantages. Troublesome impurities are also formed, in particular because the reaction is carried out in the presence of acetic anhydride. The latter also reacts with the amino acids and a certain amount of N-acetylated amino acid is then always formed, which product is difficult to separate from the N-formylated amino acid.
Another formulation process has been disclosed in U.S. Pat. No. 4,789,757. It consists in reacting the amino acid with a very large excess of formamide. Thus, in the majority of examples, a molar amount of formamide 10 times greater than that of the amino acid is used. N-Formylaspartic acid, N-formylglycine, N-formylalanine and N-formylphenylalanine are thus prepared. However, the methods for isolating these N-formylated derivatives from the reaction mixture and for recovering them with the minimum possible in the way of impurities are not disclosed.
For some applications, such as applications in the pharmaceutical or food field, it is necessary to have available an N-formylleucine of high purity. If the formylation of leucine is carried out by the process described above by means of formamide, the N-formylleucine obtained will certainly not be contaminated by impurities such as N-acetylated derivatives but it will generally still comprise unreacted leucine, isoleucine and other impurities present in the starting material, and the reaction products of these impurities with formamide.
The subject-matter of the invention is consequently a process for producing N-formylleucine having a purity at least equal to 98%.
The process according to the invention consists in preparing N-formylleucine by reaction of leucine with formamide and is characterized in that, when the reaction is complete, the N-formylleucine is precipitated, at a temperature of approximately 0° to 40° C., by mixing the reaction medium with water and an acid, so that the final pH of the mixture is approximately 2 to 3.
It has also been found that the process is improved if the reaction of leucine with formamide is carried out in the presence of formic acid and in particular in the presence of approximately 0.1 to 2 mol of formic acid per mole of leucine.
The process of the invention makes it possible to obtain N-formylleucine with the desired purity, at least equal to 98%, and with a good yield, generally of the order of 80 to 90%.
The leucine which is used as starting compound is a known compound which is available commercially in the D, L and DL forms. It generally comprises isoleucine as the main impurity.
The N-formylation reaction can be carried out as shown in U.S. Pat. No. 4,789,757 with a considerable excess of formamide, generally with approximately 10 mol of formamide per mole of amino acid.
However, it has now been found that it is preferable not to use a very large excess of formamide and that an amount of formamide of less than 4 mol per mole of leucine and in particular of: approximately 1.5 to 3.5 mol per mole of leucine is highly suitable.
It is also advantageous to carry out the N-formylation reaction in the presence of formic acid. Use is preferably made of approximately 0.5 to 1 mol of formic acid per mole of leucine.
The reaction is carried out by heating the mixture, composed of formamide and leucine and, optionally, formic acid, at a temperature of between approximately 80° C. and 110° C., preferably between approximately 90° C. and 100° C., and preferably under an inert atmosphere, such as under a nitrogen atmosphere. It generally lasts from 1 to several hours.
When it is complete, it is then advantageous, if an amount of formamide greater than the preferred amount has been used, to distill off a portion of the formamide remaining in the medium after the reaction, so as not to complicate the precipitation operation because of this excess of reactant.
The N-formylleucine is subsequently precipitated. To do this, use is made of a large amount of water, preferably of between approximately 1.5 and 5 parts per part by weight of starting formamide, and of an acid, in particular an inorganic acid, for example hydrochloric or sulphuric acid, so that there is a pH of approximately 2 to 3, preferably of approximately 2 to 2.5, at the end of the precipitation.
According to one alternative form, in carrying out the precipitation operation, acid is first of all added to the water needed for the precipitation, in order for its pH to be acidic, preferably of approximately 2 to 3, and then the reaction medium and the acidified water are mixed, preferably by pouring the reaction medium into the acidified water or by simultaneously pouring the reaction medium and the acidified water into a reactor. Throughout the duration of the precipitation, the mixture is preferably maintained at a pH of approximately 2 to 3 and more particularly of approximately 2 to 2.5 by adding acid to the mixture, if necessary. According to this alternative form, it is also advantageous, before carrying out the precipitation operation, to dilute the reaction mixture with water, for example in an amount of between approximately 0.2 and 0.4 part per part by weight of formamide.
According to another alternative form, the reaction medium is first of all mixed with water and then the pH of the mixture is brought to and maintained at approximately 2 to 3, preferably approximately 2 to 2.5, by addition of acid.
The acid used is more particularly sulphuric acid.
It is also important for the temperature during the precipitation to be maintained between approximately 0° C. and 40° C., preferably between approximately 5° C. and 20° C.
The N-formylleucine spontaneously crystallizes but it is also possible to introduce a seed in order to promote the crystallization.
On completion of the crystallization, the N-formylleucine crystals are recovered using one of the usual methods. The crystals obtained in particular have a size sufficient to be easily filtered off, washed and dried.
The dry N-formylleucine yield is generally greater than 80%.
The purity of the N-formylleucine is equal to or greater than 98%, often even greater than 99%.
The N-formylleucine obtained according to the process of the invention can be used directly as intermediate in carrying out syntheses of peptides or in preparing pharmaceutical products.


REFERENCES:
patent: 4789757 (1988-12-01), Carter
patent: 0 080 119 A1 (1983-06-01), None

LandOfFree

Say what you really think

Search LandOfFree.com for the USA inventors and patents. Rate them and share your experience with other people.

Rating

Process for producing N-formylleucine of high purity does not yet have a rating. At this time, there are no reviews or comments for this patent.

If you have personal experience with Process for producing N-formylleucine of high purity, we encourage you to share that experience with our LandOfFree.com community. Your opinion is very important and Process for producing N-formylleucine of high purity will most certainly appreciate the feedback.

Rate now

     

Profile ID: LFUS-PAI-O-2518442

  Search
All data on this website is collected from public sources. Our data reflects the most accurate information available at the time of publication.