U.S. patent application number 13/203020 was filed with the patent office on 2012-02-23 for estrus synchronization preparations & effective cidr-less protocols.
Invention is credited to Hal Witt, Jennifer Yoakum.
Application Number | 20120046518 13/203020 |
Document ID | / |
Family ID | 42356159 |
Filed Date | 2012-02-23 |
United States Patent
Application |
20120046518 |
Kind Code |
A1 |
Yoakum; Jennifer ; et
al. |
February 23, 2012 |
Estrus Synchronization Preparations & Effective CIDR-Less
Protocols
Abstract
An injectable preparation of estrogenic and progestrogenic
hormonal compounds in an anhydrous excipient injected to
synchronize estrus in non-menstruating placental female animals,
and a five to nine day protocol to apply the unique preparation to
achieve estrus synchronization.
Inventors: |
Yoakum; Jennifer; (Liberty
City, TX) ; Witt; Hal; (Longview, TX) |
Family ID: |
42356159 |
Appl. No.: |
13/203020 |
Filed: |
January 26, 2010 |
PCT Filed: |
January 26, 2010 |
PCT NO: |
PCT/US10/00199 |
371 Date: |
August 24, 2011 |
Related U.S. Patent Documents
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Application
Number |
Filing Date |
Patent Number |
|
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61206077 |
Jan 26, 2009 |
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Current U.S.
Class: |
600/35 ;
514/170 |
Current CPC
Class: |
A61K 31/567 20130101;
A61K 31/57 20130101; A61K 45/06 20130101; A61K 31/565 20130101;
A61K 31/565 20130101; A61K 2300/00 20130101; A61K 2300/00 20130101;
A61K 2300/00 20130101; A61K 47/14 20130101; A61K 31/573 20130101;
A61K 2300/00 20130101; A61P 15/08 20180101; A61K 31/573 20130101;
A61K 31/567 20130101; A61K 47/10 20130101; A61K 31/57 20130101;
A61K 9/0019 20130101 |
Class at
Publication: |
600/35 ;
514/170 |
International
Class: |
A61D 19/02 20060101
A61D019/02; A61P 15/08 20060101 A61P015/08; A61K 31/57 20060101
A61K031/57 |
Claims
1. An injectable preparation, administered by intramuscular
injection, for promoting CIDR-less estrus synchronization in
healthy, mature, cycling female cattle, comprising: (a) a steroidal
estrogenic compound such as, for example, estradiol 17-.beta.,
estradiol benzoate, or any derivatives, analogs, or agonists
thereof, and any combinations thereof, in an amount effective to
induce estrus in healthy, mature, cycling female cattle; and (b) a
steroidal progestin such as, for example, progesterone,
hydroxyprogesterone, medroxyprogesterone, altrenogest, norgestomet,
levonorgestrel, or other progestogenic compound or any derivatives,
analogs, or agonists thereof, and any combinations thereof, for
preserving the corpus luteum intact for a period of at least five
days after its injection; and (c) an anhydrous excipient compounded
with said estrogenic compound and said progestin to produce a
compound suitable for intramuscular injection.
2. A preparation as in claim 1, wherein said progestogenic
concentrations are at least 30 mg/mL after compounding.
3. A preparation as claimed in claim 1, wherein said estrogenic
compound, said progestin, and said excipient are combined in
quantities such that the said estrogenic compound is in
concentrations of at least 0.025% by volume relative to the overall
injectable preparation.
4. A preparation as claimed in claim 1, wherein the said estrogenic
compound and the said progestin are combined in volumes such that
the said estrogenic compound is in concentrations in the range of
from 0.05 to 1.0% by volume relative to the overall volume of the
injectable preparation.
5. A preparation as claimed in claim 1, wherein the said estrogenic
compound and the said progestin are combined in volumes such that
the said estrogenic compound is in concentrations in the range of
from 0.06 to 0.125% by volume relative to the overall volume of the
injectable preparation.
6. A CIDR.RTM.-less method for improving estrus synchronization and
fertilization rates in one or more healthy, mature, cycling, female
cattle that are at least 45 days postpartum--without utilizing an
intravaginal hormone dispensing device--where the method comprises
the sequential steps of: (a) administering, on Day Zero, by
intramuscular injection to each of the healthy, mature, cycling,
female cattle a first steroidal hormone preparation comprising a
combination of an estrogenic compound and a progestin compounded in
an anhydrous excipient, to induce estrus in healthy, mature,
cycling, female cattle; and (b) thereafter, sometime on Day X
(where Day X is from 5 to 8 days after Day Zero), administering a
second preparation comprising prostaglandin, or its biologically
functional equivalent, to each of the healthy, mature, cycling,
female cattle in an amount effective to initiate estrus; and (c)
thereafter, from 12 to 36 hours after administering said second
preparation, administering to each of the healthy, mature, cycling,
female cattle a third preparation comprising an estrogenic compound
in an amount to force ovulation.
7. The method of claim 6, further comprising the additional step of
attempting to impregnate each of the healthy, mature, cycling,
female cattle on Day Z not less than twelve hours after
administering said third preparation.
8. The method of claim 7 wherein said first steroidal hormone
preparation comprising a combination of an estrogenic compound and
a progestin to induce estrus in healthy, mature, cycling, female
cattle is administered within the first twelve hours on Day Zero,
said second preparation is administered within four hours of 8:00
am on Day X where Day X is 6 to 7 days after Day Zero, the said
third preparation is administered within four hours of 1:00 pm on
Day Y, and the said attempting to impregnate step comprises
artificial insemination within four hours of 2:00 pm on Day Z.
9. The method of claim 8 wherein said first steroidal hormone
preparation comprising a combination of an estrogenic compound and
a progestin to induce estrus in healthy, mature, cycling, female
cattle is administered within the first twelve hours on Day Zero,
said second preparation is administered within one hour of 9:00 am
on Day X where Day X is 6 days after Day Zero, the said third
preparation is administered within one hour of 1:00 pm on Day Y,
and the said insemination is performed within one hour of 2:00 pm
on Day Z.
10. The method of claim 6 wherein the said first steroidal hormone
preparation is an injectable preparation, administered by
intramuscular injection, for promoting CIDR-less estrus
synchronization in healthy, mature, cycling female cattle,
comprising: (a) a steroidal estrogenic compound such as, for
example, estradiol 17-.beta., estradiol benzoate, or any
derivatives, analogs, or agonists thereof, and any combinations
thereof, in an amount effective to induce estrus in healthy,
mature, cycling female cattle; and (b) a steroidal progestin such
as, for example, progesterone, hydroxyprogesterone,
medroxyprogesterone, altrenogest, norgestomet, levonorgestrel, or
other progestogenic compound or any derivatives, analogs, or
agonists thereof, and any combinations thereof, for preserving the
corpus luteum intact for a period of at least five days after its
injection; and (c) an excipient compounded with said estrogenic
compound and said progestin to produce a compound suitable for
intramuscular injection.
11. The method of claim 6 wherein the said first steroidal hormone
preparation is an injectable preparation, administered by
intramuscular injection, for promoting CIDR-less estrus
synchronization in healthy, mature, cycling female cattle,
comprising: (a) a steroidal estrogenic compound such as, for
example, estradiol 17-.beta., estradiol benzoate, or any
derivatives, analogs, or agonists thereof, and any combinations
thereof, in an amount effective to induce estrus in healthy,
mature, cycling female cattle; and (b) a steroidal progestin such
as, for example, progesterone, hydroxyprogesterone,
medroxyprogesterone, altrenogest, norgestomet, levonorgestrel, or
other progestogenic compound or any derivatives, analogs, or
agonists thereof, and any combinations thereof, for preserving the
corpus luteum intact for a period of at least five days after its
injection; and (c) an anhydrous excipient compounded with said
estrogenic compound and said progestin to produce a compound
suitable for intramuscular injection, and further, wherein said
estrogenic compound, said progestin, and said excipient are
combined in quantities such that the said estrogenic compound is in
concentrations in the range of from 0.05% to 1.0% by volume
relative to the overall injectable preparation.
12. The method of claim 7 wherein the said first steroidal hormone
preparation is an injectable preparation, administered by
intramuscular injection, for promoting CIDR-less estrus
synchronization in healthy, mature, cycling female cattle,
comprising: (a) a steroidal estrogenic compound such as, for
example, estradiol 17-.beta., estradiol benzoate, or any
derivatives, analogs, or agonists thereof, and any combinations
thereof, in an amount effective to induce estrus in healthy,
mature, cycling female cattle; and (b) a steroidal progestin such
as, for example, progesterone, hydroxyprogesterone,
medroxyprogesterone, altrenogest, norgestomet, levonorgestrel, or
other progestogenic compound or any derivatives, analogs, or
agonists thereof, and any combinations thereof, for preserving the
corpus luteum intact for a period of at least five days after its
injection; and (c) an excipient compounded with said estrogenic
compound and said progestin to produce a compound suitable for
intramuscular injection, and further, wherein the said estrogenic
compound and the said progestin are combined in volumes such that
the said estrogenic compound is in concentrations in the range of
from 0.06 to 0.125% by volume relative to the overall volume of the
injectable preparation.
13. A method for improving estrus synchronization and fertilization
rates in one or more healthy, virgin heifers--without utilizing an
intravaginal hormone dispensing device--where the method comprises
the sequential steps of: (a) administering, on Day Zero, by
intramuscular injection to each of the healthy, virgin heifers a
first steroidal hormone preparation comprising a combination of an
estrogenic compound and a progestin to induce estrus in healthy,
virgin heifers; and (b) thereafter, sometime on Day X (where Day X
is 5 to 8 days after Day Zero), administering a second preparation
comprising prostaglandin, or its equivalent, to each of the
healthy, virgin heifers in an amount effective to initiate estrus;
and (c) thereafter, sometime on Day Y (where Day Y is one day later
than Day X) but not less than 12 hours after administering said
second preparation, administering to each of the healthy, virgin
heifers a third preparation comprising an estrogenic compound in an
amount to force ovulation; and (d) thereafter, sometime on Day Z
(where Day Z is one day later than Day Y) but not less than 12
hours after administering said third preparation, inseminating each
of the healthy, virgin heifers.
14. The method of claim 13 wherein said first steroidal hormone
preparation comprising a combination of an estrogenic compound and
a progestin to induce estrus in healthy, virgin heifers is
administered within the first twelve hours on Day Zero, said second
preparation is administered within four hours of 8:00 am on Day X
where Day X is 6 to 7 days after Day Zero, the said third
preparation is administered within four hours of 1:00 pm on Day Y,
and the said insemination is performed within four hours of 2:00 pm
on Day Z.
15. The method of claim 14 wherein said first steroidal hormone
preparation comprising a combination of an estrogenic compound and
a progestin to induce estrus in healthy, virgin heifers is
administered within the first twelve hours on Day Zero, said second
preparation is administered within one hour of 9:00 am on Day X
where Day X is 6 days after Day Zero, the said third preparation is
administered within one hour of 1:00 pm on Day Y, and the said
insemination is performed within one hour of 2:00 pm on Day Z.
Description
CROSS-REFERENCE & PRIORITY CLAIM TO A PROVISIONAL
APPLICATION
[0001] This application claims the benefit under 35 U.S.C.
A.sctn.119(e) of U.S. Provisional Application 61/206,077, filed
Jan. 26, 2009, entitled "Estrus Synchronization Compounds and
Protocols". That provisional application, in its entirety, is
incorporated by reference into this application.
BACKGROUND OF THE INVENTION
[0002] A. Technical Field.
[0003] The present invention generally relates to the fields of
artificial insemination and embryo transfer in animals that undergo
estrus and, more particularly, to the protocols and pharmaceuticals
used to facilitate, manage, and control estrus synchronization in
such animals.
[0004] B. Estrus Synchronization.
[0005] In animal production, "estrus synchronization" traditionally
refers to artificially synchronizing the estrous cycle throughout a
group of female animals using hormone manipulation and supporting
protocols. The animals may include such mammals as ruminants or
non-menstruating placental females such as female cattle.
[0006] Cattle producers seek estrus synchronization in their herds
because of its many economic benefits. First, estrus
synchronization can reduce the number of days after a cow calves
before she can begin a new pregnancy. This shortened interval
increases the number of offspring a cow can produce in her
lifetime. Second, and perhaps most importantly, because any
particular breeding period is of limited duration (e.g., 45 to 90
days), and yet it may take 18 to 21 days to confirm a pregnancy
after insemination, it is tremendously valuable to confirm
pregnancy for as much of the herd as possible, as soon as possible,
preferably with the first attempt at the start of the breeding
period. If the entire herd can be estrus synchronized in the first
week of the breeding period, then there is usually ample
opportunity for a second and third opportunity to conceive during
the remainder of the breeding period, while those who do not
achieve estrus early will have fewer opportunities. Cows that do
not conceive during the regular breeding period require additional
interventions at the producer's expense. Third, cows that conceive
early in the breeding season will produce calves that weigh more at
weaning because the calves are older. Fourth, cows that calve early
will have more days postpartum before the beginning of the next
breeding season, which serves to improve the cow's overall health.
Fifth, estrus synchronization also allows the producer to
synchronize the feed, supplements, immunizations, and treatments
for all the cows in the herd, which allows for more efficient
management and greater opportunities for profit. Additionally,
estrus synchronization allows producers to breed virgin heifers
three weeks before inseminating the main herd, which gives the
heifers additional recovery time after their first calving before
the next breeding season begins.
[0007] For these and other benefits, producers attempt to
coordinate the onset of estrus in their animals, which, without
intervention, is typically asynchronous across a group of animals.
To achieve a synchronized onset of estrus, producers use hormones
and supporting protocols to intervene in their animals' estrous
cycles.
C. Estrous Cycle.
[0008] The estrous cycle comprises recurring physiologic changes
that are induced by mammalian reproductive hormones. The mammalian
reproductive system includes the hypothalamic system which releases
gonadotropin releasing hormone (GnRH), as well as the pituitary
system that secretes follicle stimulating hormone (FSH) and
luteinizing hormone (LH). The mammalian ovaries release hormones
that include estrogens and progesterone. Species that have estrous
cycles characterized by non-menstruating placental females reabsorb
the endometrium if fertilization and conception does not occur
during that particular cycle. In such species, females are
generally only sexually active during the estrus phase of their
cycle, a condition commonly referred to as being "in heat."
Therefore, an animal may be described as "in estrus" when it is in
that particular part of "the estrous cycle."
[0009] There are four basic phases to the estrous cycle in mammals
and, therefore, in all female cattle (including genus Bos of the
taurine and zebu species), ewes, does, cows, nannies, mares, and
female ruminants. In the initial phase, the proestrus phase, one or
more follicles of the ovary begin to grow. Typically this phase can
be as short as one day, or as long as three weeks, depending upon
the mammalian species. Under the natural influence of estrogen, the
lining of the uterus (the endometrium) develops. The second phase
is estrus proper and refers to the days when the animal is "in
heat." In this phase, under regulation by gonadotropic hormones,
ovarian follicles mature and estrogen secretions begin to influence
the reproductive system. In female cattle, ovulation may occur
spontaneously during this phase. Estrus may continue for a number
of days, followed by interestrus. During the third phase,
metestrus, the indications of estrogen stimulation diminish and the
corpus luteum begins to form. The corpus luteum is essential for
establishing and maintaining a pregnancy. The corpus luteum
produces progesterone which: (a) prepares the uterus for pregnancy
by thickening and developing the endometrium, (b) maintains the
pregnancy if fertilization occurs, and (c) inhibits the cattle from
showing signs of standing estrus and ovulating. Generally, the
larger the corpus luteum the more progesterone it produces, and in
the presence of progesterone, the dominant follicle will not
rupture, meaning no ovulation occurs. Further, the corpus luteum
secretions inhibit LH and FSH. Also, the uterine lining begins to
secrete progesterone. This third phase is generally short and may
last one to five days. The final stage of the estrous cycle is
diestrus, which is characterized by the production of progesterone
by the corpus luteum. In the absence of a pregnancy, the diestrus
phase terminates with the regression of the corpus luteum. The
lining in the uterus is not shed (as in menstruating mammalian
species) but is reconfigured for the next cycle.
D. Current Techniques for Estrus Synchronization.
[0010] Today, beyond true synchronization of estrus among a herd,
estrus synchronization also encompasses a wide variety of hormonal
manipulation techniques that attempt to control the date(s) on
which a particular female can either be successfully inseminated or
otherwise impregnated. One of the most reliable synchronization
techniques uses a controlled intravaginal drug release (CIDR.RTM.)
device that is inserted vaginally and left in place to introduce
hormones into the animal. CIDR.RTM. devices are marketed under the
CIDR.RTM. trademark by InterAg of Hamilton, New Zealand, although
we use the term in our descriptions to refer generically to such
vaginal inserts.
[0011] FIGS. 1 and 2 show two typical protocols for using the
CIDR.RTM. product. There are many different protocols that require
a CIDR.RTM. product; the two examples shown in FIGS. 1 and 2 are
not an exhaustive list. FIG. 1 (Prior Art) depicts a CIDR.RTM.
protocol that requires heat detection before artificial
insemination (AI).
[0012] FIG. 2 (Prior Art) depicts a CIDR.RTM. protocol with timed
artificial insemination (TAI) not dependent upon detection of
estrus.
[0013] CIDR.RTM. cattle inserts typically contain 1.38 grams of
progesterone in elastic rubber molded over a nylon spine. The
devices are typically packaged in a plastic pouch and are inserted
using a special applicator device. CIDR.RTM. cattle inserts are
administered intravaginally, one per animal, in beef cows and
heifers. The CIDR.RTM. cattle insert releases progesterone during a
seven day treatment period. CIDRs.RTM. typically allow for
sustained release of hormones to delay estrus for five to fourteen
days while the cow's corpus luteum is hopefully held intact to
allow proper development of the cow's follicles until full estrus
is allowed to proceed, often under the influence of a prostaglandin
injection.
[0014] Recommended administration, in order to assure satisfactory
synchronization, includes an injection of Lutalyse.RTM. sterile
solution (dinoprost tromethamine) that is typically given to all
synchronized cattle either at the time of, or within one day
before, CIDR.RTM. removal. Removal of the insert on treatment Day
Six or Seven results in a drop in plasma progesterone, ideally
triggering estrus within three days.
[0015] Referring again to FIG. 1 (Prior Art), the use of a
CIDR.RTM. insert is shown to extend from Day Zero to Day Seven of
the protocol. Associated with placement of the insert on Day Zero
is the administration of an injectable gonadotropin releasing
hormone (GnRH). On Day Six or Seven, with removal of the CIDR.RTM.
insert, prostaglandin (such as Lutalyse.RTM.) is typically
administered by injection. In the protocol shown in FIG. 1, the
user must investigate and detect symptoms of heat in the animal,
and thereafter, carry out the artificial insemination (AI) process.
This heat detection and AI process typically occurs between Day
Seven and Day Thirteen as calculated from Day Zero, the day of the
placement of the CIDR.RTM. device.
[0016] FIG. 2 (Prior Art) depicts another CIDR.RTM. protocol that
is similar in most respects to the protocol described in FIG. 1.
However, rather than rely upon the detection of heat in the animal,
this second protocol calls for a time period of 60-66 hours after
Day Seven (and the removal of the CIDR.RTM. insert) for a further
administration of a dose of GnRH and subsequent AI. As indicated in
FIG. 2, this typically results in a Day Ten artificial insemination
of the animal.
[0017] Besides CIDR.RTM.-based synchronization protocols, there are
also "CIDR.RTM.-less" synchronization protocols--meaning methods
that do not require the use of a CIDR.RTM. or any intravaginal
device to administer hormones.
E. Problems with Existing Protocols.
[0018] To date, many different hormonal preparations and related
protocols are well known to those of ordinary skill in the art.
However, the success rates associated with the prior art--with or
without a CIDR.RTM.--remain less than desirable, typically allowing
for successful implantation or insemination in fewer than half of
the attempts.
[0019] Additionally, various problems are known to exist with the
use of the CIDR.RTM. insert. Although CIDR.RTM. promoters assert
much better results, it is not unusual to experience conception
rates as low as 20% with many CIDR.RTM. protocols, and even lower
rates with virgin heifers. Moreover, results are diminished by high
rates of infection and vaginitis, as well as animal discomfort and
stress that are fairly common with CIDR.RTM. inserts. Use of the
CIDR.RTM. insert requires careful cleaning of the vulva to avoid
infection, and many protocols still require the detection of heat
for effective AI. It would be desirable, therefore, to establish a
reliable estrus synchronization protocol that does not require the
use of a CIDR.RTM. insert.
SUMMARY OF THE INVENTION
[0020] The present invention includes both unique preparations and
protocols that, among other benefits, (1) dramatically improve
estrus synchronization rates in healthy, mature, cycling female
cattle as well as in virgin heifers, (2) help preserve the corpus
luteum, (3) improve the success rate of embryo transfer
(transplantation), and (4) enable CIDR.RTM.-less estrus
synchronization, thereby avoiding the infections, vaginitis, and
animal discomfort that occur with CIDR devices.
[0021] The present invention is embodied in an injectable
preparation for promoting CIDR-less estrus synchronization in
non-menstruating placental females, which includes: (a) a steroidal
estrogenic compound such as estradiol 17-.beta. (E2), estradiol
benzoate, or the like in an amount effective to induce estrus in
healthy, mature, cycling female cattle; and (b) a steroidal
progestin such as progesterone, hydroxyprogesterone,
medroxyprogesterone, altrenogest, norgestomet, levonorgestrel, or
other progestogenic compound or any derivatives, analogs, or
agonists thereof, and any combinations thereof, for preserving the
corpus luteum intact for a period of at least five days after its
injection; and (c) an excipient compounded with an estrogenic
compound and a progestin to produce an injectable compound.
[0022] One embodiment of the preparation is made where the
estrogenic compound, the progestin, and the excipient are combined
in quantities such that the estrogenic compound is in
concentrations from 0.05% to 1.0% by volume relative to the overall
injectable preparation, preferably from 0.06 to 0.125%, and the
combination is in a form suitable for intramuscular delivery.
[0023] Another embodiment of the present invention includes a
preparation which, when compounded, obtains progestogenic
concentrations of at least 30 mg/mL.
[0024] The present invention is also embodied in protocols for
improving estrus synchronization and fertilization rates in
non-menstruating female animals without utilizing an intravaginal
hormone dispensing device. The protocols are based on four ordered
steps: (1) injecting intramuscularly one or more healthy, cycling
females with a first steroidal hormone preparation to prepare such
females to induce estrus; (2) administering a second preparation
principally prostaglandin, or its biologically functional, to
actually initiate estrus; (3) administering a third preparation of
an estrogenic compound to force ovulation; and (4) impregnating
each female animal, either through artificial insemination or
otherwise.
[0025] The first preparation is administered by intramuscular
injection within the first twelve hours of Day Zero. The second
preparation is administered by intramuscular injection no sooner
than the beginning of Day Five but no later than the end of Day
Eight. The third preparation is administered about one day after
the second preparation, and the animals are artificially
inseminated or naturally bred about one day after the
administration of the third preparation.
[0026] The first step is to administer to each female animal by
intramuscular injection approximately 2 cm.sup.3 of a first
steroidal hormone preparation combining an estrogenic compound with
a progestin. Next, 5 to 8, preferably 6 or 7, and ideally 6 days
later, each female receives an intramuscular injection of a second
preparation, namely prostaglandin, or its biologically functional
equivalent, in an amount to initiate estrus and ovulation. The
final step in the typical process is the further step of
impregnating the female animals, preferably through artificial
insemination. The two injections properly administered, combined
with a subsequent estradiol injection, improve estrus
synchronization rates, increase conception rates in virgin heifers,
and achieve synchronization without the need for a CIDR.RTM., which
omission reduces the risk of vaginitis and vaginal infections and
decreases animal discomfort.
[0027] A related preparation and protocol is involved in an
alternative embodiment that is used to synchronize donor cows. Such
embodiments include a base protocol the DonorSync.TM. Protocol, to
use the preparation of the foregoing embodiments to prepare donor
cows for embryo transfer (ET). In ET practice, donor cows are often
treated with a hormone protocol to facilitate embryo production.
The donor cow is inseminated at the appropriate time, and the
embryos are collected about 6-8 days after breeding. When the donor
cow's embryos are collected, they need to be immediately
transplanted into recipient cows. For the embryo to survive and
become a live pregnancy, each recipient cow's estrous cycle must be
in the same state as the donor cow's cycle so that the recipient
cow's uterus is prepared to receive the embryo. The present
invention protocol, using DonorSync.TM., with its higher
concentration of progestin than in the SuperSync.TM. preparation,
has been shown to be highly affective in resetting the follicular
wave. When followed by an FSH and PG protocol, the DonorSync.TM.
yields excellent results in producing transferable embryos.
[0028] Further, the present invention's protocols and preparations
are also used in ET using frozen embryos.
[0029] It will also be appreciated that many invaluable aspects of
the present invention could also be understood or expressed from
other vantage points. From the context of a kit, for example, it
can be appreciated that aspects of the present invention allow for
a kit that contains both preparations and protocols to be followed
to achieve estrus synchronization. Still other embodiments of the
invention relate to products made by the described protocols as
well as apparatuses and systems for performing all or part of such
protocols.
[0030] Many other objects, features, and advantages of the present
invention will be evident from the remainder of this application in
light of a more exhaustive understanding of the numerous
difficulties and challenges faced by the prior art, which in turn
will be evident to those skilled in the art. In the end, while
there are many alternative variations, modifications, and
substitutions within the scope of the invention, one of ordinary
skill in the art should consider the scope of the invention from a
review of the claims appended hereto (including any amendments made
to those claims in the course of prosecuting this and related
applications) as considered in the context of the prior art and the
various descriptions of this application.
DETAILED DESCRIPTION OF PREFERRED EMBODIMENTS
[0031] The teachings of the present inventions are preferably
embodied as injectable hormonal preparations and as protocols that
use such preparations in order to improve estrus synchronization
and achieve a much higher percentage of estrus synchronization as
well as the desired preservation of larger corpus luteum sizes. It
will be understood by those skilled in the art that the protocols
and administered preparations described below in conjunction with
the present invention are further facilitated by other techniques
known to optimize the benefits of estrus synchronization. These
other techniques include the maintenance of adequate nutrition as
well as efforts to maintain sufficient overall body scores for the
cows and heifers. Those skilled in the art will also recognize
alternative applications of the compounds and methods of the
present invention that may include the preparation of recipient
cows for implantation of embryos.
[0032] For reference in various aspects of the preferred
embodiments of the present invention, Applicant has developed a
preferred formulation for a steroidal hormonal preparation that
Applicant expects to commercialize under the designation
"SuperSync.TM.." The formulation of the SuperSync.TM. preparation
is a combination of an estrogenic compound, a progestin, and
appropriate excipients to achieve an injectable form. By an
"estrogenic" compound, we mean an artificial or synthetic estrogen
(i.e., a steroidal estrogenic compound) or derivatives, analogs, or
agonists thereof, and any combinations thereof. Likewise, by a
"progestin," we mean an artificial or synthetic progesterone (i.e.,
a steroidal progestogenic compound), such as, for example,
progesterone, hydroxyprogesterone, medroxyprogesterone,
altrenogest, norgestomet, levonorgestrel, or other progestogenic
compound, or derivatives, analogs, or agonists thereof, and any
combinations thereof. Preferably, the estrogenic compound is
selected from the group of estradiol 17-.beta. or estradiol
benzoate, or their derivatives, analogs, agonists and the like.
[0033] To achieve a preparation that is suitable for intramuscular
injection, the estrogenic compound and the progestin are compounded
with an excipient that preferably combines an anhydrous carrier
base (such as sesame seed oil, cotton seed oil, olive oil) with one
or more excipient solvents. In the preferred embodiments, the
excipient solvents are phenylmethanol (approximately 10% of the
final volume) and a co-solvent benzoic acid phenylmethyl ester
(approximately 10% of the final volume). The excimer is compounded
with at least 1 mg/mL estrogenic compound (preferably 1.25 mg/mL).
For a 2 cm.sup.3 dose, preferred preparations provide from 2 to 3
mg estradiol 17-.beta., which is an amount effective to initiate
estrus or, in other words, to reset the follicular wave in a heifer
or a cow (including a non-bovine cow), and at least 30 mg/mL
progestogenic compound (preferably more than 40 mg/mL and
approximately 60 mg/mL), which has been found to be effective at
preserving the corpus luteum intact for five or more days from the
date of injection. The maximum amount of estradiol 17-.beta. should
not ever exceed 5 mg per dose.
[0034] FIG. 3 depicts the methodology of a SuperSync.TM. Protocol
according to the processes of the present invention. FIG. 4
provides a flow chart describing in a step-by-step manner the
methodology of the protocol of the present invention. Initially
(Step 402 in FIG. 4), it is necessary to identify a Day Zero for
the protocol by determining a time period of 45-60 days postpartum.
Starting the protocol no earlier than this time period ensures an
involuted uterus. On Day Zero (Step 404), 2 cm.sup.3 of the
proprietary formulation (SuperSync.TM.) is administered by
intramuscular (IM) injection, preferably in the neck of the animal.
The formulation of the proprietary preparation may vary moderately,
but in the preferred embodiment includes progesterone and estradiol
in a ratio of 60 mg/mL progesterone to 1.25 mg/mL estradiol. This
proprietary preparation should be administered in the first twelve
hours of Day Zero.
[0035] The administered preparations described above in the
protocol (SuperSync.TM. Protocol) generally carry out the following
functions in the process of estrus synchronization. The proprietary
preparation of progestin and estradiol provides a unique
simultaneous combination of effects. As indicated above, the
progestin component keeps a cow or heifer out of heat and extends
the estrous cycle. The progestin/estradiol combination, when
administered at Day Zero, resets the follicular wave in the animal.
The prostaglandin (PG) prepares for initiation of heat in the
animal. Estradiol, administered subsequent to prostaglandin,
further facilitates ovulation. Gonadotropin releasing hormone
(GnRH), as indicated above, is a hormone that triggers ovulation or
starts development of a new follicular wave. Lutenizing hormone
(LH) also triggers ovulation and follicle stimulating hormone (FSH)
promotes follicular formation. Progestin mimics natural
progesterone produced by the corpus luteum after ovulation which
prepares the uterus for pregnancy and serves to keep the cow or
heifer from coming back into heat.
[0036] Estradiol is the most common, and generally considered to be
the most effective estrogen hormone. Estradiol 17-.beta. is a
naturally occurring hormone that tends to result in the quickest
reaction in cows. Estradiol benzoate is a possible alternative to
estradiol 17-.beta.. The compositions contained in the proprietary
preparation (SuperSync.TM.) administered on Day Zero, therefore,
carry out the important functions towards the goals of the
protocol. The progestin helps to preserve the corpus luteum in good
condition by shutting down the pituitary function. In essence the
progestin ensures that the corpus luteum remains intact. The
estradiol functions to reset the follicular wave. Specifically,
estradiol 17-.beta. (E2) knocks off the dominant follicle, thereby
releasing the ovacyte (ovulation), which becomes the egg (once
fertilized). The ruptured follicle also either becomes the corpus
luteum or adds to it (presuming that a corpus luteum was
pre-existing).
[0037] The second action step of the SuperSync.TM. Protocol (Step
406) takes place on Day X, where Day X is any day from 5 to 8 days
after Day Zero, although it may start as soon as the corpus luteum
has adequately redeveloped. On Day X, therefore, 5 cm.sup.3 of
prostaglandin (PG) is administered by intramuscular injection,
preferably in the neck of the animal, and preferably within four
hours of 8:00 am and optimally within one hour of 9:00 am. In the
preferred embodiments, the prostaglandin (PG) utilized may be a
product marketed as Lutalyse.RTM. (a Registered Trademark of
Pharmacia & Upjohn Co. of Michigan) which contains the
naturally occurring prostaglandin F2.alpha. (dinoprost) as the
tromethamine salt. Each milliliter contains dinoprost tromethamine
equivalent to 5 mg dinoprost. Alternately, in the preferred
embodiments, the prostaglandin (PG) may be a product marketed as
Estrumate.RTM. (a Registered Trademark of Schering-Plough of New
Jersey) which is a synthetic prostaglandin analogue structurally
related to prostaglandin F2.alpha.. Each milliliter of the
colorless, aqueous solution contains 263 mcg of cloprostenol sodium
(equivalent to 250 mcg of cloprostenol). In any case, the
prostaglandin functions to put a heifer or cow into heat, thereby
disrupting the yellow tissue that makes up the corpus luteum,
causing the release of scent and other signs of heat. This process
thereby forces ovulation which is the start of complete estrus.
[0038] The third action step (Step 408) typically occurs on Day Y,
which is the day immediately following Day X, but should occur
within 12 to 36 hours after administering the second preparation
wherein 1 cm.sup.3 of estradiol is administered, sometime on Day Y,
preferably within four hours of 1:00 pm, and optimally within one
hour of 1:00 pm. This administration of estradiol functions to
stimulate the pituitary to release hormones causing ovulation
(i.e., ovacyte to drop, vulva to swell, etc.). The estradiol used
here and elsewhere in the preferred protocols is preferably
estradiol 17-.beta. (E2) or estradiol benzoate, although it should
be appreciated that various other natural or synthetic estrogenic
compounds, or their derivatives, analogs, or agonists, and any
combinations thereof, may be used as less preferred alternatives.
It will also be appreciated that, at the risk of marginalizing the
benefits of the dosing in the preferred embodiments, alternate
concentrations and/or volumes may also be used. For instance, it is
appreciated that certain aspects of the invention can be
accomplished through administration of other concentrations of the
estrogen compound, so long as at least one milligram of estradiol
17-.beta., or the biologically functional equivalent dose of an
alternate steroidal estrogenic compound, is administered.
[0039] The final action step (Step 410) in the SuperSync.TM.
Protocol is carried out on Day Z, which is the day immediately
following Day Y, wherein insemination is effected on Day Z,
preferably within four hours of 2:00 pm, and optimally within one
hour of 2:00 pm. Insemination may be carried out with frozen or
thawed semen, with actual fertilization typically occurring 4-6
hours after insemination.
[0040] Those skilled in the art will recognize that the protocol of
the present invention as described above is less labor intensive
than those protocols associated with the use of the CIDR.RTM.
device and may be used on female animals that are genetically
ill-suited to CIDR.RTM. use (e.g., Beefmaster cows that typically
cannot retain a CIDR.RTM. in place) or individual females that do
not tolerate the CIDR.RTM. (e.g., vaginal scarring from previous
pregnancies). The present invention's protocol is less traumatic
for the cows and heifers, and may be expected to generally maintain
better overall health during administration of the protocol than
with the CIDR.RTM. system. Although the protocol and the
proprietary preparation have been described in conjunction with a
set of preferred embodiments, it will be recognized by those of
ordinary skill in the art that certain modifications to the
protocols and to the formulation may be made without departing from
the spirit and scope of the invention. Variations based on the size
of the herd and/or the size and overall health of the individual
cows and heifers, will become apparent. Likewise, variations on the
ratio of the compounds making up the proprietary preparation will
be apparent to those skilled in the art upon a consideration of the
specific applications to which the protocol is directed. In other
words, some such variations would be appropriate for use of the
protocols in conjunction with natural or artificial insemination
versus a similar use of the protocols for embryo implantation. As
indicated, these variations, given the basic concepts of the
present invention, do not necessarily depart from the spirit and
scope of the invention.
[0041] The results that Applicants are able to achieve using the
present invention are surprisingly better than the prior art. For
instance, although still unpublished, controlled studies have
indicated dramatically greater conception rates in virgin dairy
Holstein heifers using a CIDR.RTM.-less protocol where the second
preparation was administered on Day 6. Similarly, although results
may vary with poor controls, estrus synchronization using the
present invention has been preliminarily reported to such a degree
that more then 90% of a herd reached estrus almost simultaneously,
reportedly with two hours of each other.
[0042] Many other objects, features, and advantages of the present
invention will be evident from the remainder of this application in
light of a more exhaustive understanding of the numerous
difficulties and challenges faced by the prior art, which in turn
will be evident to those skilled in the art. In the end, while
there are many alternative variations, modifications, and
substitutions within the scope of the invention, one of ordinary
skill in the art should consider the scope of the invention from a
review of the claims appended hereto (including any amendments made
to those claims in the course of prosecuting this and related
applications) as considered in the context of the prior art and the
various descriptions of this application.
[0043] For instance, an alternative embodiment of the present
invention involves use of a form of the first preparation (and in a
higher dose) in donor cows to achieve successful estrus
synchronization in preparation for embryo production, collection,
and transfer into recipient cows. This embodiment is effective in
taurine cows but is especially effective in Brahman or other zebu
cows. FIG. 5 depicts the methodology of a DonorSync.TM. Protocol
according to the processes of the present invention. For this
embodiment, one or more of the preferred variations of the Day Zero
preparation from the previous embodiments is used to prepare the
donor cow. The injected amount of such preparations, however, is
increased--preferably from 30% to 150% more than in the prior
description. Most preferably, the donor cow is injected with the
first preparation in a dose of from 3.0 to 4.5 cm.sup.3 and,
beginning four days later, a multi-day course of
follicle-stimulating hormone (FSH) administered intramuscularly
twice daily to produce more ovacytes, concluding with injections of
prostaglandin to rupture all of the developed follicles. After
ovulation, the donor cow is inseminated, usually several times in
over a 12 to 24 hour period. Thereafter, on Day T, typically Day 7,
the embryos are collected from the donors and transferred either to
the recipients or preserved by freezing.
* * * * *