U.S. patent application number 10/624997 was filed with the patent office on 2004-03-18 for purification process.
Invention is credited to Fuenfschilling, Peter, Schenkel, Berthold.
Application Number | 20040050782 10/624997 |
Document ID | / |
Family ID | 10799753 |
Filed Date | 2004-03-18 |
United States Patent
Application |
20040050782 |
Kind Code |
A1 |
Fuenfschilling, Peter ; et
al. |
March 18, 2004 |
Purification process
Abstract
This invention provides a process for purifying a cyclosporin,
e.g. cyclosporin A, or a macrolide, to a high degree of purity on a
large scale. In another aspect this invention provides a bulk
quantity of cyclosporin A with an impurity level of less than about
0.7%, e.g. about 0.5%, and compositions thereof.
Inventors: |
Fuenfschilling, Peter;
(Allschwil, CH) ; Schenkel, Berthold; (Basel,
CH) |
Correspondence
Address: |
THOMAS HOXIE
NOVARTIS, CORPORATE INTELLECTUAL PROPERTY
ONE HEALTH PLAZA 430/2
EAST HANOVER
NJ
07936-1080
US
|
Family ID: |
10799753 |
Appl. No.: |
10/624997 |
Filed: |
July 23, 2003 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
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10624997 |
Jul 23, 2003 |
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10021117 |
Oct 29, 2001 |
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6620325 |
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10021117 |
Oct 29, 2001 |
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09652295 |
Aug 31, 2000 |
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09652295 |
Aug 31, 2000 |
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09271672 |
Mar 18, 1999 |
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09271672 |
Mar 18, 1999 |
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08926722 |
Sep 10, 1997 |
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Current U.S.
Class: |
210/634 ;
210/806; 422/256 |
Current CPC
Class: |
C07D 259/00 20130101;
A61K 38/00 20130101; C07K 7/645 20130101 |
Class at
Publication: |
210/634 ;
210/806; 422/256 |
International
Class: |
B01D 011/00 |
Foreign Application Data
Date |
Code |
Application Number |
Sep 11, 1996 |
GB |
9618952.7 |
Claims
1. A process for purifying on a large scale a product from a
feedstock containing one or more impurities having closely-related
physical properties to the product, which process comprises feeding
the feedstock into an extraction column under conditions adapted
for separating more- or less-polar impurities from the feedstock,
wherein a lighter phase flows counter to a heavier phase, thereby
forming an output in one phase containing the product containing
less more- or less-polar impurities so that the output contains the
product in a substantially purified form, and wherein the lighter
phase comprises heptane and acetone or heptane and isopropanol, the
heavier phase comprises water and acetone or water and isopropanol,
and the product is a cyclosporin, a rapamycin or an ascomycin.
2. A process as claimed in claim 1, wherein the lighter phase
comprises about 25 wt-% n-heptane and about 75 wt-% acetone, or
about 90 wt-% n-heptane and about 10 wt-% isopropanol.
3. A process as claimed in claim 1, wherein the heavier phase
comprises about 50 wt-% water and about 50 wt-% acetone, or about
68 wt-% water and about 32 wt-% isopropanol.
4. A process for purifying on a large scale a product from a
feedstock containing one or more impurities having closely-related
physical properties to the product, which process comprises the
steps of a) feeding the feedstock into a first extraction column
under conditions adapted for separating more- or less-polar
impurities from the feedstock, wherein a lighter phase flows
counter to a heavier phase, thereby forming a first output in one
phase containing the product containing less more- or less-polar
impurities, and b) feeding the first output into a second
extraction column under conditions adapted for separating less- or
more-polar impurities respectively from the first output, wherein
the lighter phase flows counter to the heavier phase, thereby
forming in one phase a second output, so that the second output
contains the product in a substantially purified form, wherein the
lighter phase comprises heptane and acetone or heptane and
isopropanol, the heavier phase comprises water and acetone or water
and isopropanol, and the product is a cyclosporin, a rapamycin or
an ascomycin.
5. A process as claimed in claim 4, wherein the lighter phase
comprises about 25 wt-% n-heptane and about 75 wt-% acetone, or
about 90 wt-% n-heptane and about 10 wt-% isopropanol.
6. A process as claimed in claim 4, wherein the heavier phase
comprises about 50 wt-% water and about 50 wt-% acetone, or about
68 wt-% water and about 32 wt-% isopropanol.
7. A process as claimed in claim 1 or claim 4, wherein the product
is Cyclosporin A, Cyclosporin D or a derivative thereof,
Cyclosporin G or a derivative thereof, rapamycin,
40-O-(2-hydroxy)ethyl rapamycin, ascomycin,
33-epi-chloro-33-desoxyascomycin, or FK506.
8. A countercurrent extraction column having between 100 and 200
compartments, and an overall efficiency of about 10 to 30%.
9. A bulk quantity of cyclosporin A with an impurity level of less
than 0.5% by area using HPLC.
10. A composition comprising as active agent cyclosporin A as
claimed in claim 9.
Description
[0001] The present invention relates to a purification process, and
in particular to a process for purifying a product from impurities
having relatively close distribution coefficients one with
another.
[0002] Extraction processes are well-known for separating
hydrocarbons in the petrochemical industry. It is known that
distribution coefficients of the hydrocarbons between given phase
systems in such applications differ substantially one from
another.
[0003] Reviews of extraction methods are published, for example, in
Kirk-Othmer Encylopedia of Chemical Technology, 4th edition, Vol.
10, p. 125-180, pub. John Wiley & Sons 1993, and in The
Handbook of Solvent Extraction, ed. Lo, Baird & Hanson, pub.
Krieger 1991. The contents of these publications are incorporated
herein by reference.
[0004] Presently large-scale purification of products of
biotechnological processes is generally carried out by extraction
or adsorption chromatography. Such products include peptides,
macrolides and proteins which are generally produced as mixtures of
products having closely-related structural and/or physical
properties. Fractional extraction processes known in the
petrochemical industry have hitherto not been applied in the
biotechnology industry on a large scale for purification of active
agents, e.g. peptides, having closely-related physical properties
one with another. The present applicants consider a major problem
to be that the respective distribution coefficients of
biotechnological products and impurities between given phases are
close one with another.
[0005] Accordingly, in one aspect, the present invention provides a
process for purifying a product from a feedstock containing one or
more impurities having closely-related physical properties to the
product, which process comprises
[0006] feeding the feedstock into an extraction column under
conditions adapted for separating more- or less-polar impurities
from the feedstock, wherein a lighter phase flows counter to a
heavier phase, thereby forming an output in one phase containing
the product containing less more- or less-polar impurities so that
the output contains the product in a substantially purified
form.
[0007] The feedstock for the above process may be provided by
output from a chromatographic purification or other
pre-purification step, e.g. decantation. This process may be
conducted such that the output serves as input for a subsequent
chromatographic purification, e.g. in a series arrangement.
[0008] In another aspect, this invention provides a process for
purifying a product from a feedstock containing one or more
impurities having closely-related physical properties to the
product, which process comprises
[0009] a) feeding the feedstock into a first extraction column
under conditions adapted for separating more- or less-polar
impurities from the feedstock, wherein a lighter phase flows
counter to a heavier phase, thereby forming a first output in one
phase containing the product containing less more- or less-polar
impurities, and
[0010] b) feeding the first output into a second extraction column
under conditions adapted for separating less- or more-polar
impurities respectively from the first output, wherein the lighter
phase flows counter to the heavier phase, thereby forming in one
phase a second output, so that the second output contains the
product in a substantially purified form.
[0011] The feedstock may be prepared by known methods, for example
by fermentation. When the product is produced in a fermentation
broth, the broth may be filtered and mixed with a solvent from
which the product may be precipitated in an impure state, e.g.
containing about 15% to about 30% by weight impurities. Typically
the fermentation broth may undergo several cleaning and work-up
steps prior to use as feedstock in the process of this invention.
Initial filtration(s) and precipitation(s) serve to remove, for
example, natural dyes and easily separable impurities from the
product, and may serve to enhance phase separation in the
extraction steps.
[0012] As used herein, the term "phase" is understood to mean a
system having at least one component. The phase may comprise a
single solvent or a mixture of, for example 2, 3 or more
solvents.
[0013] As used herein, the term "closely-related physical
properties" is understood to mean that the product is difficult to
separate from the one or more impurities. The closely-related
physical properties may include, for example, the respective
distribution coefficients of the product and one or more impurities
between two phases. For a cyclopeptide, e.g. a cyclosporin, and in
particular cyclosporin A, ratios of distribution coefficients, i.e.
distribution coefficient of cyclosporin A/distribution coefficient
of impurity, may be between about 3 and about 0.4, e.g. between
about 1.5 and about 0.8. These ratios are known as
selectivities.
[0014] As used herein, the term "large scale" as applied to
purification plant, is understood to mean a plant having an output
of about one or more tonnes of purified product per annum, e.g. 10
tonnes per annum, or more e.g. around 20 to around 40 tonnes.
[0015] As used herein, the term "impurity" is understood to mean an
undesirable component.
[0016] The process of this invention may be applied to a wide
variety of products, e.g. peptides, such as cyclosporins, for
example Cyclosporin A and derivatives thereof, Cyclosporin D and
derivatives thereof, or Cyclosporin G and derivatives thereof. An
example of a Cyclosporin D derivative is, for example
([3'-desoxy-3'-oxo-MeBmt].sup.1-[Val].sup.2-Ci- closporin) as
disclosed in EP 296122; or macrolides, for example rapamycins and
derivatives thereof, and ascomycins and derivatives thereof,
produced for example by fermentation. Examples of macrolides which
may be purified using the process of this invention include
rapamycin; 40-O-(2-hydroxy)ethyl rapamycin as described in
PCT/EP93/02604; ascomycin; 33-epi-chloro-33-desoxyascomycin as
described in EP 427680 in Example 66a; ascomycin derivatives
disclosed e.g. in EP 569337 and in EP 626385, for example
5,6-dehydro-ascomycin as disclosed in EP 626385; or an ascomycin
derivative known as FK506.
[0017] A method for producing Cyclosporin A is disclosed, for
example, in Example 1 of British patent specification 1,491,509.
Methods of preparing FK506 are described in EP 184162.
[0018] A variety of columns are known and available commercially.
In one embodiment of this invention, a single column is used which
is a countercurrent column adapted for mechanical agitation and/or
stirring of the feedstock/phase system. In another embodiment of
this invention, two or more columns are used, at least one of which
is a countercurrent column adapted for mechanical agitation and/or
stirring of the feedstock/phase system.
[0019] Preferably the column(s) includes mechanical agitation, e.g.
rotary agitation or reciprocating plate. An extraction column
adapted for mechanical agitation is available commercially from the
Kuehni company, Switzerland.
[0020] In another aspect, this invention provides a countercurrent
liquid-liquid extraction column adapted for rotary agitation having
a sufficient number of trays or compartments to effect, in use,
separation of a pharmaceutical from impurities.
[0021] In a further aspect, this invention provides the use of a
countercurrent liquid-liquid extraction column adapted for rotary
agitation for separating a pharmaceutical from impurities.
[0022] The column(s) may be adapted for temperature adjustment of
the phases. A jacket may be provided, for example, to maintain a
desired temperature within the column. When purifying, for example
a cyclosporin, temperatures may be maintained within each column at
between about 0 and about 100.degree. C., preferably between about
20 and about 80.degree. C., and more preferably between about 30
and about 60.degree. C.
[0023] It may be advantageous to conduct the process using a first
column in series arrangement with a second and a third column,
wherein the second column is in parallel arrangement with the third
column. The Applicants contemplate such a configuration when, for
example, the plant is to be situated in a building having a
covering of limited height.
[0024] The phases chosen for the extraction step(s) of the process
may depend on the distribution coefficient between phases, and
solubility of the product in each phase under consideration. Fast
and efficient separation of the phases is desirable, and separation
may be substantially complete, for example within about one minute,
preferably 30 seconds, more preferably within about 20 seconds.
[0025] A two-phase system is used in the process of this invention,
wherein the phases are immiscible or substantially immiscible one
with another. At least one phase may be an aqueous phase. The
two-phase system separates to form a lighter, e.g. non-aqueous
organic, phase comprising at least one solvent component, and a
heavier, e.g. aqueous, phase comprising at least one solvent
component and water. The product to be purified may be more readily
soluble in one phase component which, therefore, serves as
extracting component.
[0026] Preferably one phase is an aqueous phase and one phase is a
non-aqueous phase. The aqueous phase may contain at least about
20%, e.g. more than about 40% by weight water, and may contain up
to 100% by weight water. The aqueous phase may be the heavier
phase.
[0027] The phase systems used in the processes of this invention
may comprise, as organic phase, at least one hydrocarbon, e.g. a
C.sub.5 to C.sub.12 alkane, e.g. n-heptane, cyclohexane or
methylcyclohexane. As aqueous phase, the present applicants
contemplate, in addition to water, ketones, e.g. acetone; esters,
e.g. amyl acetate, n-butyl acetate or isopropyl acetate; or
alcohols, e.g. methanol, ethanol, propanol, isopropanol, n-butanol,
s-butanol, t-butanol or a pentanol. Acetone is a preferred
extracting solvent for cyclosporin A.
[0028] It will be appreciated that at least one component is common
to both phases. The common component may comprise any of the
above-mentioned solvents, e.g. water; hydrocarbons; alcohols;
ketones; or esters.
[0029] The present applicants have found an n-heptane/acetone/water
system to be effective for purifying acomycin and derivatives
thereof, rapamycin and derivatives thereof and cyclosporin A. In
one embodiment, the following phase system is used to purify
cyclosporin A: the lighter phase comprises a mixture in weight-% of
about 75% n-heptane and about 25% acetone; the heavier phase
comprises a mixture of about 50% acetone and about 50% water. The
water content of the lighter, substantially non-aqueous phase is
typically less than 10% by weight, preferably less than about 2% by
weight based on the total weight of the non-aqueous phase.
[0030] In another embodiment, the following phase system is used to
purify 40-O-(2-hydroxy)ethyl rapamycin: the lighter phase may
comprise a mixture in weight-% of about 75%, e.g. 74%, n-heptane,
about 25%, e.g. 25.5% acetone and a small quantity of water, e.g.
0.5% water. The heavier phase may comprise a mixture of a small
amount e.g. 0.4% n-heptane, about 50%, e.g. 52.8%, acetone and
about 50%, e.g. 46.8%, water.
[0031] The present applicants have found an
n-heptane/isopropanol/water system to be effective for purifying
33-epi-chloro-33-desoxyascomycin. In one embodiment, the following
phase system is used to purify 33-epi-chloro-33-desoxyascomycin:
the lighter phase comprises a mixture in weight-% of about 90%
n-heptane, e.g. 89%, and about 10% isopropanol, e.g. 11%; the
heavier hase comprises a mixture of about 32% isopropanol, e.g.
31%, and about 68% water, e.g. 69%.
[0032] The distribution coefficient is defined herein as the ratio
of the concentration (in g/unit volume) of product in the lighter
phase to the concentration (in g/unit volume) of product in the
heavier aqueous phase at 40.degree. C. The following distribution
coefficients are obtained for cyclosporins between 75%
n-heptane/25% acetone as lighter organic phase, and 50% acetone/50%
water as heavier aqueous phase:
1 Cyclosporin Distribution coefficient (to one decimal place) A 0.8
B 0.4 C 0.3 D 1.7 Dihydro-A 1.0 G 1.3 L 0.5 U 0.6
[0033] From the above, it will be apparent that the respective
distribution coefficients for cyclosporins are relatively close one
with another. When purifying Cyclosporin A, for example, the
impurities to be separated from Cyclosporin A typically include
other cyclosporins.
[0034] One advantage of this process is that Cyclosporin A may be
prepared substantially free from impurities such as other
Cyclosporins, e.g. Cyclosporin B, Cyclosporin C, or
dihydro-Cyclosporin A. Thus the respective amounts of cyclosporin
derivative impurities, e.g. Cyclosporin B, Cyclosporin C and
Cyclosporin G in Cyclosporin A purified using the process of this
invention, are typically at or below analytically detectable limits
using HPLC. Thus the impurities in cyclosporin A are found to
amount to less than about 0.7% by area using HPLC, e.g. about 0.5%
or less, e.g. about 0.3%.
[0035] In another aspect, therefore, this invention provides a bulk
quantity, i.e. 1 kg or more, e.g. 10 kg or more, e.g. 20, 30, 40,
50 kg or more cyclosporin A with an impurity level of less than
about 0.7%, e.g. 0.5% or less, by area using HPLC. This invention
further provides a bulk quantity of cyclosporin A having a purity
level of 99.5% or greater, e.g. 99.7% or greater. The impurities
found in purified cyclosporin A of this invention consist of other
cyclosporins.
[0036] In another aspect this invention provides a composition
comprising cyclosporin A as active agent in a bulk purified form,
wherein the amount of impurities present is below 0.7%, e.g. 0.5%
by area using HPLC.
[0037] The composition may be orally administrable and may be an
emulsion, microemulsion, a microemulsion preconcentrate or a solid
dispersion. The composition may comprise components disclosed for
example in published UK patent application GB 2 222 770, the
contents of which are incorporated herein by reference.
[0038] Distribution coefficients for 40-O-(2-hydroxy)ethyl
rapamycin are found to lie between 0.7 and 1.4 when using
n-heptane/acetone/water phases. Ratios of distribution
coefficients, i.e. selectivities, for rapamycin to
40-O-(2-hydroxy)ethyl rapamycin typically range from about 1.4 to
about 2.3, e.g. around 1.5.
[0039] The distribution coefficient for
33-epi-chloro-33-desoxyascomycin is found to be around 7.6 using
the above n-heptane/isopropanol/water phase system. The selectivity
of ascomycin derivative side product to
33-epi-chloro-33-desoxyascomycin is found to be approximately
1.44.
[0040] The pH of the aqueous phase, when present, is typically
between about 2 and about 9, for example pH 7.
[0041] The extraction process conditions, including the number of
theoretical stages, may be selected using routine experimentation
and, for example, computer simulation. A program suitable for such
a purpose is available commercially under the trade name ASPEN
PLUS.
[0042] Exact conditions for working this invention typically depend
on a number of factors, including the dimensions of the column(s),
fluid dynamics, stirrer speed, efficiency and number of trays or
compartments.
[0043] Compartments having an efficiency of about 10 to 30%, e.g.
12 to 25%, may be suitable for the column(s) of this invention.
Column efficiency, as used herein, is understood to mean the number
of actual stages divided by the number of theoretical stages.
[0044] It will be appreciated by those skilled in this art that an
extraction column, in use, may exhibit a flooding point. The
respective flow rates of the phases may therefore be chosen to
maintain the system in the or each column at below the flooding
point, for example around 10 or 20% below the flooding point. In
this way, high turbulence and fast mass transfer between phases may
be obtained.
[0045] The present applicants have found that for a given column,
the flooding point may be reached when a particular stirrer speed
is reached and/or exceeded, hereinafter referred to as the critical
speed. Stirrer speeds applied in a column in the process of this
invention may be between about 15 and about 200 rpm. On a large
scale, typical stirrer speeds may be between about 5 and about 60
rpm. The applicants have obtained good results using a stirrer
speed about 10 to 20% below the critical speed.
[0046] In one embodiment of this invention, the same solvent
system(s) may be used in each column, and the respective flow rates
may be varied so that the desired product may be extracted into
either phase. The respective flow rates may be determined using
routine experimentation and computer simulation using, for example,
the above-mentioned computer program.
[0047] The processes of this invention may result in a highly pure
product, in particular when combined, for example, with
distillation, precipitation and/or crystallisation steps. After a
crystallisation step, solvent(s) may be distilled off, collected
and recycled. The product may be obtained in known solid form.
[0048] The process of this invention may be applied to the
purification of, for example, cyclosporins to achieve purity
greater than 98.5%, e.g. 99.3% or greater by area using HPLC, and
improved overall yield compared to chromatographic
purification.
[0049] The order of extraction used in the two-step process of this
invention may be reversed: the less polar impurities may be removed
from the impure product in the first column, and the more polar
impurities removed in the second column. If the product to be
purified is to be separated from a single impurity, or only from
less polar components, or only from more polar components, the
process may be adapted to include a single extraction step, for
example, only above step a) or only step b).
[0050] Following is a description by way of example only and
illustrated by accompanying drawings, of a purification process of
this invention conducted in a pilot plant having an output capacity
of about 1 kg/day purified product, and on a commercial scale
having an output capacity of at least about 10 kg/day purified
product.
[0051] FIGS. 1 to 4 each show a chromatograph of Cyclosporin A and
impurities at stages of the purification process.
[0052] FIG. 5 is a schematic representation of phase flows.
[0053] Cyclosporin A is produced in a fermentation broth which is
worked up and filtered. FIG. 1 shows a chromatograph of the
filtered product containing Cyclosporin A; Cyclosporin A is
represented by the peak at 24.418.
[0054] Pilot Plant Scale
[0055] The filtered product is mixed with acetone, and Cyclosporin
A is partially crystallised from the acetone to form a feedstock
F1. Feedstock F1 is fed into a first extraction column (1, pilot
scale) half way up the column at a central zone f1. Column 1 is
fitted with mechanical agitation. A lighter (organic) phase LP,
consisting of 25 wt-% acetone and 75 wt-% n-heptane, is fed into
column 1 at a lower zone b1. A heavier (aqueous) phase HP,
consisting of 50% water and 50% acetone, is fed into column 1 at an
upper zone al. The following flow rates are used in the first
column:
2 litres/hour Feed (F1) 4.2 heavier (aqueous) phase (HP) 12.7
lighter phase (LP) 13
[0056] A product stream S1 containing Cyclosporin A with more polar
impurities, exits from the lower zone in the aqueous phase at d1;
less polar impurities are removed in the lighter phase P1 which
exits from column 1 at upper zone c1. FIG. 2 shows a chromatograph
of the stream S1 at this stage; Cyclosporin A is represented by the
peak at 26.248.
[0057] Product stream S1 serves as feedstock F2 for a second column
(2, pilot scale), and is fed into column 2 halfway up column 2 at
central zone f2. The second column is also fitted with mechanical
agitation. The lighter phase LP is fed into column 2 at a lower
zone b2, and heavier (aqueous) phase HP is fed into column 2 at
upper zone a2. More polar impurities are removed in the heavier
phase P2 which exits from column 2 at lower zone d2. Cyclosporin A
is removed in product stream S2 with lighter phase from column 2 at
upper zone c2. FIG. 3 is a chromatograph of the stream S2 mixture
at this stage; Cyclosporin A is represented by the peak at
26.598.
[0058] Arrowheads in FIG. 5 indicate directions of phase/product
stream flows with respect to the columns.
3 The following flow rates are used in the second column:
litres/hour Feed (F2) 3.1 heavier (aqueous) phase (HP) 9.7 lighter
phase (LP) 17.7
[0059] Organic solvents are distilled from the Cyclosporin
A/lighter phase mixture which is subsequently mixed with acetone,
from which Cyclosporin A is crystallised. Cyclosporin A at a purity
of >98.5% is obtained, as determined by high-performance liquid
chromatography. FIG. 4 shows a chromatograph with purified
Cyclosporin A represented by the peak at 26.368.
[0060] The internal diameter of each column employed in the pilot
plant is 6 cm.
[0061] Commercial Scale
[0062] A. For large, commercial, scale operation, using similar
geometric parameters and hydrodynamic conditions to those used in
the pilot plant, the Applicants calculate that 40 theoretical
stages are required; the first column has an overall 25% efficiency
and 160 compartments. The first column has a height of about 25 m
and an internal diameter of about 30 cm. For the second column, the
Applicants calculate that 30 theoretical stages are required, and
120 compartments. The second column is calculated to have a height
of about 25 m and an internal diameter of about 80 cm.
[0063] A purification plant is constructed using the above
parameters with the two columns in series arrangement.
[0064] A bulk quantity of 50 kg cyclosporin A is produced. On
analysis of the cyclosporin using HPLC, impurities amount to less
than 0.5% by area. This implies a purity of at least 99.5% by
weight.
[0065] B. Using similar geometries and hydrodynamic conditions to
those used in the pilot extraction column, the Applicants calculate
that 44 theoretical stages at an efficiency of approximately 27%
are required in the first CCE step. The first column is calculated
to have a height of approximately 27 m, an internal diameter of
approximately 45 cm and 160 compartments. For the second column,
the Applicants calculate that 62 theoretical stages at an
efficiency of approximately 38% are required. The second column is
calculated to have a height of about 34 m and an internal diameter
of about 70 cm, and 160 compartments.
[0066] A purification plant is constructed using these parameters.
A third column is provided substantially identical in construction
and dimensions with the second column. The first column is in
series arrangement with the second and third columns. The second
and third columns are in mutually parallel arrangement. Output from
the first column is divided and fed into the second and third
columns.
[0067] A bulk quantity of 100 kg cyclosporin A is produced. On
analysis of the cyclosporin using HPLC, impurities amount to less
than 0.5% by area. This suggests a cyclosporin A purity of at least
99.5% by weight.
[0068] The present invention provides a continuous process for
purifying on a large scale a product(s) from impurities, which
product(s) and impurities have similar physical properties one with
another. Greater purities at a particular yield, or greater yields
at a particular purity, may be achieved using this process than by
using chromatographic purification. The process is more economical
in operation since product streams do not have to be divided and/or
recycled as is generally required with absorption
chromatography.
[0069] The countercurrent process of this invention is surprisingly
more efficient than purification by chromatography, and multiple
recycling steps are avoided. The process of this invention allows
simulation and adjustment of process parameters more readily in
order to achieve a desired purity than when using chromatography.
Compounds of closely-related structure are suprisingly more easily
separated one from another on a commercial scale, e.g. Cyclosporin
A from Cyclosporins B and C.
* * * * *