Note: Descriptions are shown in the official language in which they were submitted.
z
600-7089
CYCLOSPORIN OPE~T~LNIC COMPOSITIO~S
The present invention relaees to novel ophthalmic compositions, i.e.
pharmaceutical compositions for admini~tration to the eye9 comprising
a cyclosporin as active ingredient and suitable for the treatment of
diseases and conditions of the eye and surrounding areas
The cyclosporins comprise a large and recognised class of peptide
compounds having pharmaceutical utility, ~or example
immunosuppressant, anti-inflammatory, and/or anti-parasitic activity
and/or activity in abrogating tumor resistance to antineoplastic or
cytostatic drug therapy. The cyclosporins include, for example,
naturally occurring fungal metabolites, such as the cyclosporins A, B,
C, D and G, as well as a wide variety of synthetic and semi-synthetic
cyclosporins, for example the dihydro- and iso-cyclosporins (see e.g.
US Patents Nos. 4,108,985; 4,210,581 and 4,220,641), l(D)-Ser]e-
-Ciclosporin (see USP 4,384,996), lO-acetyl-(D)Ser~8-Ciclosporin (see
USP 4,764,503), [~-fluoro-(D~Ala]~-Ciclosporin (see UK Patent
Application 2,206,119A), lVal~2-[(D)methylthio-Sar]3- and
[Dihydro-MeBmt]l-[Val]2-[(D)methylthio-Sarl3-Ciclosporin [see USP
4,703,033] and very many more.
Of the cyclosporins, the most widely investigated to date is
cyclosporin A, also known and referred to hereinafter as Ciclosporin
and commercially available under the Registered Trade Mark SANDIMMUN
or SANDIMMUNE. Ciclosporin has been shown to suppress selectively a
variety of T-lymphocyte functions, including prevention of maturation
and expression of sensitized T-lymphocytes in cell mediated immune
responses, and is now successfullq and widely used in the suppression
of organ tran~plant rejection. Ciclosporin has also been used
systemically in the treatment of intraocular inflammatory or
autoimmune diseases, such as uveitis. However, because of the side
5~
- 2 - 600-7089
effects associated with systemic therapy, Ciclosporin has had only
limited use in treating such cond~tions of the eye.
Effective topical administr~tion of Ciclosporin to the eye would
reduce or eliminate to a large extent the systemic side ~ffects by
restricting activity to ehe locus of the condition being treated and
proposals to this effect have been made, (see e.g. USP ~,649,047).
However, utility and effectiveness of Ciclosporin in treating diseases
and conditions of the eye has remained hindered by the lack of a
suitable composition which is acceptable or effective for topical use.
A composition is required, which does not cau~e patient discomfort and
which permits a convenient administration regimen9 that is, does not
require unduly frequent administration, while providing adequate drug
substance delivery both to the external and, in particular, the
internal regions of the eye.
Similar considerations apply to other cyclosporins known from the art,
e.g. proposed for use as immunosuppressants or anti-inflammatory
agents, for example cyclosporin G, also known and referred to
hereinafter as lNva]2-Ciclosporin.
The present invention provides novel ophthalmic compositions
comprising a cyclosporin as active lngredient which meet difficulties
hitherto encountered in the art, e.g. as described above. The
compositions of the invention are intended for topical application,
i.e. for application to or at the surface of the eye, e.g. to the
cornea or corneal epithelium, or to the immediate areas surrounding
the eye, for example the inner surfaces of the upper or lower lid.
Applied topically as aforesaid, the compositions of the invention are
useful for the treatment of diseases or conditions of the eye or of
the surrounding or associated organs or tissues, for example the tear
glands and ducts, especially immune mediated or inflammatory diseases
or conditions. More especially the compositions of the invention are
useful for the treatment of diseases or conditions ~hich involve
undesirably elevated immuno-response or inflammatory reaction or event
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- 3 - 600-7089
as a component or part of their etiology, in particular autoimmune
disPases of the eye. Diseases and conditions which may be treated with
the compositions of the invention include, for example uveitis (both
anterior and posterior), chronic ker~titis, conjunctivitis (including
in particular vernal conjunctivltis), vernal keratocon~unctivitis,
keratoconjunctivitis sicca, and keratoplasty (i.e. use in relation to
corneal transplant), as well as inflammatory or immune mediated
conditions induced by ocular surgery in general. The compositions of
the invention may also be employed for the induction or maintainance
of tearing, for example where tear-function is impaired, e.g. in
consequence of any disease or condition as aforesaid.
The ophthalmic compositions of the invention are surprisingly found to
cause little or no irritation or patlent discomfort and to provide a
therapeutic effece at convenient application rates ln the corneal and
internal regions of the eye, including the anterior chamber, posterior
chamber, vitreous body, aqueous humor, vitreous humor, cornea,
iris/ciliary, lens, choroid/retina, or sclera or in surrounding organs
or tissues of the eye, for example the tear duct or tear gland.
~ore particularly the present invention provides in a first aspect:
A) An ophthalmic composition comprising a cyclosporin as active
ingredient and comprising (1) an ophthalmically acceptable
vegetable oil and (2) an ophthalmically acceptable petroleum jelly
as carrier medium.
The compositions of the invention are intended for topical ophthalmic
application, i.e. application to the surface of the eye, e.g. to the
cornea or corneal epithelium, or to the immediate surrounding regions
of the eye.
By "ophthalmically acceptable" is meant appropriate or allowable for
administration to the eye, e.g. sa~e fcr topical ophthalmic
application at dosages to be administered.
- 2~S~
- 4 - 600-7089
Preferred cyclosporins for use in the compositions of th~ inv~ntion
are cyclosporins having immunosuppressant or anti-inflammatory
activity, e.g. as hereinbefore described, in particular Cicloqporin. A
further preferred cyclosporin for use in the compositions of the
invention is [Nva~2-Ciclosporin. The compositions of the invention are
suitably in the form of an ointment with the defined components (1
and ~2) comprising the ointment base.
The cyclosporin, e.g. Ciclosporin, is suitably present in the
compositions of the invantion in an amount of from about 0.01 to abou~
lG%. (Unless otherwise indicated, all percentages herein and in the
accompanying claims are by weight based on the total weight of the
composition.) Preferably the cyclosporin is present in an amount of
from about 0.05 to about 10%, especially from about 0.05 to about 5%,
more preferably from about 0.1 to about 2.5%. Most preferably the
cyclosporin is present in an amount of about 0.1, 0~5, 1.0 or 2.0%.
Component (1) [vegetable oil] is suitably present in the compositions
of the invention in an amount of at least 25~. Suitably component (1)
is present in an amount of from about 25 to about 65~, preferably from
about 25 to about 45~, more preferably from about 35 to about 45~, and
especially about 40 to about 45X.
Component (2) [petroleum iellY] is suitably present in the
compositions of the invention in an amoun~ of at least 25~, preferably
at least 50X. Suitably component (2) is present in an amount of from
about 25 to about 65~, preferably from about S0 to about 65~, more
preferably from about 50 to about 60~, and especially from about 50 to
about 55%.
Components (1) and (2) are suitably present in the compositions of the
invention in a ratio of from about 1:2 to 2:1 p.p.w., e.g. about 1:1
P-p.w. .
Component (1) may comprise any appropriate vegetable oil including,
e.g. olive oil, arachis oil, corn oil, castor oil or sesame oil or
5C~2
- 5 _ 600-7089
mixtures thereof. Preferably however, component (1) comprises corn
oil, e.g. as described in the ~ANDBOOK OF P~ IACEUTICAL EXCIPIENTS
~herein "RPE"), ~oint publication of the American Pharmaceutical
Association and the Pharmaceutical Society of Great Britain, at page
91.
Component (2) may comprise any appropriate petroleum jelly product
e.g. as available under the names white pretolatum (USP 8 United
States Pharmacopoeia), white soft paraffin (BP/EP . British/European
Pharmacopoeias), white pe~roleum ~elly or petrol~um ~elly or under the
names petrolatum yellow (USP~, yellow soft paraffin (BP/EP), yellow
petrolatum or yellow petroleum ~elly - for further definition see HPE,
p.p. 194-195. Especially preferred as component (2) are white
petrolatum products, e.g. having a Color (Maximum, Lovibond Color - 2"
Cell) = ca. 0.5 to 18Y, 0.5R.
Compositions in accordance with the invention also suitably comprise
(3) an emulQifier. Suitable compon~ntY (3) include any ophthalmically
acceptable emulsifier, e.g. as known in the art. Preferred emulsifiers
for use in the compositions are non-ionic emulsifiers, in particular
non-ionic lanolin derivatives or extracts, especially lanolin alcohols
(see ~PE, p. 164) or sterol-based emulsifiers generally. Examples of
suitable emulsifiers for use in the compositions of the invention are
products such as known and commercially available under the Trade Name
Amerchol, in particular the products Amerchol 400, C, CAB, H9, L-99,
4-500 and RC and especially Amerchol CAB. lFor further defin~tion of
the above products see Fiedler, "Lexikon der Hilfstoffe, 3rd Edition
(1989), p.p. 138-139].
Components (3) when present are suitably present in the compositions
of the invention in an amount of from about 0.5 to about 10%,
preferably from about 0.5 to about 5%, more preferably from about 1 to
2.5X, e.g. about 2%.
Compositions in accordance with the invention may also include any
further components suitable for use in ophthalmic preparations, for
5~)2
- 6 - 600-7089
example preserving or anti-microbial agents. In particular they will
suitably comprise (4) a preserving agene, chlorobutanol (e.g. as
anhydrous chlorobutonal) - see HPE, p.p. 72-73 - be$ng particularly
suitable. Such additional components, e.g. components (4), when
present will suitably be present in amounts of up to about 5.0%, more
suitably up to about 2.0X, e.g. from about 0.01 to aboue 2.0% or to
about 5.0%, preferably from about 0.1 to about 1.0%, e.g. ca. 0.5X.
Compositions in accordance with the present invention may thus
suitably comprise, e.g. ~he following ingredients in the relative
indicated amounts:
CYCLOSPORIN le.g. CICLOSPORIN
OR ~Nva] 2 -CICLOSPORIN] 0.01 to 10
~ 25 to 65%
(2) _ 25 to 65~ preferably 50 to 65%
(3) 0.5 to lOX
(4) 0.1 t~ 5%
plus any further components to a total of 100%, whereby components (1)
and (2) are preferably present in a ratio of about 1:2 to 2:1 p.p.w. 9
and whereby preferred ranges for individual components may
independently be selected from those hereinbefore indicated. The
following example is illustrative of the compositions of the
invention.
~PLB
COMPONENT
CYCLOSPORIN (e.g. CICLOSPORIN) 2.0 ~
(l) CORN OIL 41.65%
(2) UHITE PETROLATUM 53.9 %
(3) NON-IONIC LANOLIN DERIVATIVES
(e.g. AMERC~OL CAB) 1.96Z
~4) C~LOROBUT~NOL (ANHYDROUS) 0.49%
TOTAL 100.0 %
o~
- 7 - 600-7089
EquiYalent compositions comprising 0.1, 0.5 and 1% cyclosporin (e.g.
Ciclosporin) may be prepared by increasing each of the components (1)
to (4) together by the required proportional amount.
The composition, which is in the form of an ointment, may be prepared
by standard technin,ues, for example, by adding (1) to (2) and heating
to about 35 to 60C to form a clear liquid, then dissolving (3) and
(4) with stirring in the clear liquid to form she vehicle for the
ointment. The Ciclosporin i9 dissolved by stirring in the 11quefied
vehicle, after which the ointment is ready for final processing.
Alternatively, the Ciclosporin can be dissolved in (l) or a portion of
(l) and the obtained solution then added eo the remaining ingredients
before further processing. For production in large quantity the
Ciclosporin is suitably dissolved in the mixture of (1) - (4) at
elevated temperature using a high torque mixer. The ob~ained solution
is then filtered asceptically and filled under sterile conditions into
containers, e.g. ointment tubes, suitably con~aining up ~o ca. Sg,
e.g. 3.5g ointment per container.
In production, compositions of the invention are preferably subjected
to filtration, e.g. asceptic filtration, on completion of dissolution
of the cyclosporin and before further processing.
Por therapeutic use, compositions of the invention will preferably be
filled into containerc appropriate, e.g. adapted, devised or intended,
for direct topical opththalmic application of the contents, for
example ointment tubes as aforesaid having a closeable outlet of
relatively small diameter and permitting ready application of the
contents to or at the surface of ~he eye, e.g. in an amount of from
about 0.05 to about 0.2 ml, e.g. of about 0.05 to about 0.1 ml.
Utility of the compositions of ~he invention may be demonstrated in
animal test models, e.g. as hereinafter described, or in clinical
trials.
2~5~;~
- 8 600-7089
TEST HET~OD
The objective of the method is to determine the penetration into and
reservoirs of radioactivity in various tissues of the rabbit eye, as
well as systemic concentrations following a single topical application
of compositions comprising tritiated Ciclosporin ("Ci-3H"), by
investigation of distribution of Ci-3H in the eye tissues and obtained
Ci-3H blood levels (systemic concentrations).
Ci-3H is prepared by introducing a tritium label at amino acid residue
1 (-MeBmt-) of Ciclosporin by standard techniques, for example, as
described in Voges et al., "Synthesis and Applications of Isotopically
Labeled Compounds", Ed. R.R. Muccino, Elsevier Press Ams~erdam,
371-376 (1986), ~n the article entitled "Tritiated Compounds for
In-Vivo Investigation". Purity and ideneity are established by NMR
spectroscopy, TLC/radio-TLC, and reverse-phase ~PLC. A 1:10 dilution
of Ci-3H with unlabeled Ciclosporin is used for testing.
For tasting, female New Zealand white rabbits ~Lab ~ab Co.) weighing 4
to 6 kilograms are used. The animals are housed in standard rabbit
cages, fed a controlled diet (Purina Chow) and allowed water ad
libitum. For approximately one hour ~ollowing application of test
composition, the rabbits are kept in a normal upright position in a
restrainer. Test compositions are administered in 20 ~1 or 19.9 mg
dosages using a calibrated pipette.
Pipettes are carefully filled so that there are no air bubbles, and
test composition is carefully expelled into the cupped inferior
cul-de-sac of both rabbit eyes. The lids are then held together gently
for one second and released. The eyes are visually monitored for any
signs of irritation, irritation induced tearing or opacity in
accordance with the Calgon modification of the Draize test using a
dissection lamp instead of a slit lamp (Draize, J.H., Appraisal for
the Safety of Chemicals in Foods, Drugs and Cosmetics, association of
Food and Drug ~fficials of the United States, Texas State Department
of Health, Austin, Texas, 1959. Modified by Calgon Consumer Products
2~s~
- 9 - 600-7089
Research Labs, Toxicology, 1973.).
After dosing, groups of 3 rabbits are sacrificPd at 0.3, 1, 2, 4, 6,
12, 24, 48 and 96 hours. Ju~t prior to sacrifioe, blood samples are
obtained from the marginal ear vein. The animals are ~hen sacrificed
by in~ection of T-61 Euthanasia Solution (~oechst) into the marginal
ear vein. Both eyes are rinsed with normal saline (0.9%NaCl) and
enucleated without rupturing surrounding blood vessels. The ocular
surface is carefully rinsed and dried with filter paper to remove any
drug remaining in the tear fluid. The aqueous humor is removed by
means of a 26 gauge 3/8 inch needle attached to a 0.2 milliliter (ml)
pipette inserted at the corneal limbus. The cornea is excised at the
limbus, following which the iris and ciliary body are dissected as a
single sample. The lens is then removed, and an aliquot of the
vitreous is collected. ~xcess vitreous i9 removed from the remaining
tissues by blotting with filter paper. The choroid-retina is scraped
from the sclera, and a sample oP the sclera is taken from the section
closest to the opt~c nerve. To avoid contamination, clean instruments
are used for the dissection of individual tissues. Each sample is
rinsed and blotted on filter paper before being weighed into
combustion cones. Blood samples (~200 ~1) are also weighed into
combustion cones. The tissue and blood samples are air-dried prior to
combustion in a Packard Tri-Carb~ Sample Oxidizer, Model 306 using
MonophaseR-40 (Packard Instrument Co.). Following combustion, the
radioactivity of the samples is determined by scintillation counting
in a Packard Tri-Carb~ Liquid Scintillation Spectrometer, ~odel 460.
To determine specific activity, a 2.0 ~1 aliquot of test composition
is dissolved in 100 ml of t.-butyl methyl ether, and 0.1 ml of the
ether solution is assayed by the combus~ion method. Using the specific
activity, the disintegrations per minute per gram (dpm/g)
concentration of radioactivity in the tissues is converted to nanogram
equivalents of Ciclosporin~gram of tissue or blood.
Radioactivity of all samples is determined within a statistical error
of 7~ at a confidence level of 95~. This implies that any net counts
lS02
- 10 - 600-7089
per minut~ (cpm) YaIue less than ~ ~ould not be significantly
diffe~ent from zero and corresponds to a detection limit of 34
picograms per gram (æg/g)-
In one series of trials employlng the above test methodology, thefollowing compositions are compared:
TEST COMPOSITION A: Composition of Example 1 ln accordance with the
present invention.
T~ST COMPOSITION B: Comparative composition, comprising Ciclosporin
and components (2) to (4) in the same amounes a~ in Example 1 and
41.~5X mineral oil in place of corn oil as component (1).
TEST COMPOSITION C: Comparative composition comprising 2% Ciclosporin
and 98~ castor oil.
(For the purposes of filling CO~POSITION B into pipettes, the
composition is first warmed to 30C to facilitate filling and then
allowed to cool to room temperature before application.)
~ESULTS
Figures 1 to 7 attached provide graphical representations for
variation in c~ncentratlon of radio~ctiv~ty In each o~ the assayed eye
tissues with time. In each graph, time is plotted along the abscissa
(horizontally) in hours up to 96 hours: concentraion is plotted along
the ordinate (vertically) in mg Eq of Ciclosporin~g of tissue, all
values being normalised to a dose of 0.335 mg Ciclosporin/eye.
Results for composition A are represented by plots with solid circles.
Results for composition B are represented by plots with solid squares.
Results for composition C are represented by plots with open inverted
triangles.
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- 11 - 500-7089
Fig. 1 provides results for the aqueous humor.
Fig. 2 provides results for the cornea.
Fig. 3 provides results for the iris/ciliary.
Fig. 4 provides results for the lens.
Fig. 5 provides results for the vitreous.
Fig. 6 provides results for the choroid/retina.
Fig. 7 provides results for the sclera.
Figs. l to 7 indicate a fairly rapid initial absorprion of Ciclosporin
with all three formulations into the anterior and posterior tissue~ of
the eye. Houever, peak concentrations in ~he cornea (Fig. 2), which
appears to serve as a reservoir for other in~raocular tissues, was
achieved ca. 3x as rapidly with composition A (ca. 2 hrs.) as compared
with composition B (ca. 6 hrs.). Moreover, concentrations achieved in
the cornea with composition A are markedly higher than with
composition B. Composition C requires ca. 12 hours to achieve maximum
concentration in the cornea.
The resultant and surprising increase in delivery to the deeper
tissues of the eye is confirmed by r~sult~ for the aqueous humor (Fig.
1) where the achieved AUC (area under curve) for composition A for the
96 hour period is ca. 2x that achieved with composition B and >3x that
achieved with composition C. [Actual AUC (96hr.) values (mg ~q/g
normalised to 0.335 mg/eye = composition A - 636: composition B - 334:
composition C - 188.] The same pattern of advantage for composition A
is maintained with respect to the iris/ciliary (Fig. 3) and the lens
(Fig. 4)
Composition A is thus seen to be markedly and surprisingly superior to
both compositons B and C with respect to delivery to anterior regions
of the eye, in particular the cornea and to the aqueous/iris/ciliary.
Compositions of the invention are thus of particular advantage in the
treatment of diseases or conditions of the eye affecting or as they
affect these regions, for example, for use in relation to corneal
transplant, to the treatment of uveitis and of kerotoconjunctivitis
sicca.
s~
- 12 - 600-70~9
As regards the posterior segmen~ of the eye, ~hile superiority of
composition A is less marked with respect to the vltreous and sclera
(Figs. 5 and 7), it is notably superior in relation to the
choroid/retina (Pig. 7), regions of the eye more typically involved in
autoimmune diseases affecting or as they affect the posterior segment.
Thus composition A provides markedly superior peak delivery as well as
superior and bet~er sustained total delivery to the choroid/retina.
Thus a recorded AUC value (96hr) in the choroid~retina (mg Eq/g
normalised to 0.335 mg/eye) for composition A is 527 as compared with
values of only 296 and 184 for compositions B and C respectively.
Composition A is thu9 further markedly and surprisingly superior to
both compositions B and C with respect to delivery to the posterior
segment of the eye, including the choroid/retina and is thus of
particular advantage in the treatment of diseases and conditions of
the eye affecting or as they affect these regions, for example for use
in the treatment of posterior uveitis as well as other immune mediated
or inflammatory retinopathies.
As will be noted from Figs. 1 to 7, comparative composition C is in
all respects markedly inferior to both compositions A and B.
~he following table provides determined data for Ciclosporin blood
level (systemic) concentration~. ~ng Eq/g normalised to 0.335 mg/eye).
L5~2
- 13 - 600-7089
TIME COMPOSITION A COMPOSITIQN B COMPOSITION C
. _ ..... . _ ._
0.50.05 + 0.04 O 0.12 + 0.12
10.97 + 1.091.36 ~ 0.560.04 ~ 0.04
21.56 i 0.710.~8 + 0.610.44 i 0.17
41.25 i 0.88 0.87 ~ 1.680.71 ~ 0.11
6Q.27 i 0.230.07 i 0.190.8~ ~ 0.48
12 O 1.30 i 3.520.46 ~ 0.27
24 O 0.57 ~ 1.5S0.63 ~ 0.50
4~ O 0.12 + 0.340.81 + 0.84
96 O O 0.18 + 0.05
.
AUC 5.96 30.68 54.49
(0-96h)
As will be seen, systemic levels measured are relatively low for all
three test compositions, but surpringly so for composition A in
comparison with both B and C. Composition A thus appear a~ markedly
superior both in t~rms of delivery to the eye and its tissues as well
as in avoidance of systemic ~nvolvement.
Yisual observation of the eye treated with composition A in the
modified Draize test showed no composition related irritation.
The advantageous the~apeutic properties of the compositions of the
invention may ~lso be demon trated ln clinical trlals, ~or example on
administrat~on to subJects exhibiting disease or conditions of the eye
as hereinabove set forth.
For the purpose of such trials, or for practical therapeutic use, e.g.
in the treatment of uveitis (anterior or posterior), vernal
conjunctivitis, vernal keratoconjunctivitis or keratoconjunctivitis
sicca, composition of the invention, e.g. the composition of example
1, are suitably admlnistered at or to the surface of the eye in
individual amounts e.g. of from ca. 0.1 to 0.2 ml from 1 to 4x daily,
e.g. depending on the particular disease or condition to be treated,
its clinical status and the effect desired. Marked improvement in
- 14 - 600-70~9
condition as compared with e.g. untreated controls are observable with
continuance of treatment, e.g. over a period o~ 1 to 2 weeks and
upwards. The compositions of the invention are found to be well
tolerated by subjects undergoing therapy, with no significant or
unto~ard irritation.
In accordance with the foregoing and in a further series of
embodiments the present invention al50 provides:
B) An ophthalmic composition as defined under (A) above in a
container appropriate for ophthalmic application of said
composition, e.g. as hereinbefore described, for example
appropriate for application of said composition to or at the
surface of the eye, e.g~ to the cornea or corneal epithelium;
C) A process for the production of an ophthalmic composition as
defined under (A) above, which process comprises bringing a
cyclosporin into intimate admixture, appropriately with the
application of warming, e.g. to a ~emperature of from about 30 to
about 60C, with component (1~ and component (2) and, optionally,
a component (3) and a component (4) as hereinbefore defined, and
optionally thereafter filling the obtained composition into an
appropriate container, e.g. as defined for (B) above;
D) A method of treating a disease or condition of the eye or of the
surrounding or aqsociated organs or tissues in a sub~ect in need
thereof, in particular of treating immune mediated or inflammatory
diseases or conditions of the eye or of the surrounding or
associated organs or tissues, which method comprises ad~inistering
a composition as defined under (A) above topically to the eye,
e.g. to or at the surface of the eye, e.g. to the cornea or
corneal epithelium; as well as
E) A composition as defined under (A) above for use in a ~ethod as
defined under (D) above, or a cyclosporin for use in the
preparation of a composition as defined under (A) above, said
5;02
- 15 - 600-7089
composition being for use in a method as defined under tD) above.
Particular sections, segments or tissues of the eye to which the
method (D) is applicable are as hereinbefore described. Particular
diseases or conditions of the eye to ~hich the method (D) is
applicable are similarly as hereinbefore described.
