KR100757265B1 - A composition for treating urinary incontinence comprising umbilical cord blood stem cells - Google Patents

Abstract

A composition for treating urinary incontinence is provided to treat urinary incontinence effectively, minimize complications after surgical operation, and reduce treatment time by applying umbilical cord blood stem cells which dose not damage to human body. A composition for treating urinary incontinence comprises umbilical cord blood stem cells including mesenchymal stem cells and mesenchymal progenitor cells and pharmaceutically acceptable carriers and is formulated as periurethral injection, wherein the umbilical cord blood stem cells are derived from human umbilical cord blood, the urinary incontinence is stress urinary incontinence, and the daily dosage of umbilical cord blood stem cells is 2.5x10^8 to 6.5x10^8 per a patient with body weight of 60kg.

Description

A composition for treating urinary incontinence comprising umbilical cord blood stem cells}

The present invention relates to a composition for treating urinary incontinence, comprising umbilical cord blood stem cells.

Urinary incontinence accounts for 10-40% of all women, and more than 200 million women worldwide suffer from urinary incontinence. Stress urinary incontinence is the most common form of female incontinence, accounting for about 50% of all incontinence. Stress urinary incontinence is a condition in which urine leaks out of the urethra due to a sudden increase in abdominal pressure, peaking in women between 45 and 49 years (approximately 65%), causing many women social and hygiene problems. Cause.

These stress urinary incontinences are divided into anatomical incontinence due to urethral hypermobiliy and intrinsic sphincter deficiency (ISD). Anatomical incontinence is performed by bladder neck suspension through the abdomen or vagina or by midurethral sling operation with tape.Intrinsic urethral sphincter dysfunction is performed by sling surgery, autologous fat, Teflon, Although urethral submucosal injections such as collagen and silicone or artificial urethral sphincter insertion are being performed, there is no perfect treatment that can be used for all urinary incontinence.

On the other hand, the stem cells are generic to the cells before fully differentiation, and can be divided into embryonic stem cells and adult stem cells. Unlike embryonic stem cells, which still have many problems in clinical application, adult stem cells are cells that have a small number of parts of the human body that have already grown, and have the ability to regenerate damaged tissues by proliferating and differentiating in case of tissue damage or stress. .

Yiou R et al. Reported the regeneration of the rhabdomyosphincter using intrinsic satellite cells, which are stem cells present in the rhabdomyophysis, after rat injury (Yiou R, Lefaucheur JP, Atala A). The regeneration process of the striated urethral sphincter involves activation of intrinsic satellite cells.Anat.Embryo. 2003; 206: 429-35), Yokoyama T, et al., Have shown that muscle-derived projenitor cells (MDPCs) are normal. Yokoyama T, Yoshimura N, Dhir R, et al: Persistence and survival autologous muscle derived cells versus bovine collagen as potential treatment of stress urinary incontinence.J Urol 2001; 165: 271-6), Tracy W, et al., Injecting allogenic muscle derived projenitor cells into nerve-cutting rat urethra to improve sphincter contractility. (Tracy W., Ji Youl Lee, George Somogyi, et al .: Improved sphincter contractility after allogenic muscle-derived progeni cel infection into the denervated rat urethra. Urology 2003; 62: 958-63).

In addition, as a clinical study conducted in humans, Strasser H et al. Performed a urethral ultrasound of autologous muscle-derived srem cells using urethral ultrasound to improve symptoms. It has been reported that the thickness of the urethra and urethral sphincter increased and the contractile force of the urethral sphincter improved (Strasser, H., Marksteiner, R., Margreiter, E., Frauscher, F., Bartsch, G., Hering, S). Stem cell therapy in treatment of urinary incontinence: first clinical result.J. Urol. 2004; 171 (Suppl.): 130.

However, a problem with conventional stem cells for the treatment of urinary incontinence is that first, stem cells must be cultured in the muscles of the old incontinence patients themselves. Although there is a huge advantage of my own stem cells, despite the claim that there is no relationship between some ages and the function of muscle stem cells, Fulle S et al. Reported that their ability to repair declines with gradual decline as they age. Fulle, S., Protasi, F., Di Tano, G., et al .: The contribution of reactive oxygen species to sarcopenia and muscle ageing.Exp. Gerontol, 2004; 39: 17-24) Renault V and Thornell LE also reported that the percentage of stem cells in skeletal muscle decreased with age (4-6% vs. 1-2%) (Renault, V., Thorne, LE, Eriksson, PO, et. al .: Regenerative potential of human skeletal muscle during aging.Aging Cell 200; 1 (2): 132-9; and Thornell, LE, Lindstrom, M., Renault, V., et al .: Satellite cell and training in the Scand.J.Med.Sci.Sports 2003; 13 (1): 48-55), Jejurikar SS, etc., showed a decrease in the capacity of cell proliferation and migration. (Jejurikar, S.S., Kuzon Jr., W.M ..: Satellite cell depletion in degenerative skeletal muscle.Apoptosis 2003; 8 (6): 573-8). After all, the cells of older people have the ability to form myotubes, but the growth rate tends to decrease compared to younger ones. If you transplant stem cells from your muscles, the effect is not enough.

In addition, a problem of conventional urinary incontinence treatment using stem cells is that the process of harvesting muscles from the arm of the patient is invasive and there is a risk of bleeding or infection at the surgical site.

Thus, the present inventors have repeated the study to improve the problem of the urinary incontinence treatment using the conventional stem cells, stem cells derived from umbilical cord blood which is a kind of adult stem cells that can be collected without causing any special harm to the human body When applied to the treatment of urinary incontinence, the present invention was found to be effective in treating urinary incontinence.

Therefore, an object of the present invention is to provide a composition for treating urinary incontinence comprising the stem cells derived from the cord blood.

According to one aspect of the invention, there is provided a composition for treating urinary incontinence comprising an umbilical cord blood stem cell and a pharmaceutically acceptable carrier. The urinary incontinence comprises stress urinary incontinence and the formulation of the composition is preferably in the form of a periurethral injection injection.

Hereinafter, the present invention will be described in detail.

The composition for treating urinary incontinence of the present invention includes umbilical cord blood stem cells.

Many clinical experiences have been reported since Gluckman E et al. Successfully performed cord cord transplantation (CBT) for the first time in Fanconi anemia (Gluckman E, Broxmeyer HE, Auerbach AD, et al. Hematopoietic reconsitution in a patient with Fanconi's anemia by means of umbilical cord blood from an HLA-identical sibling.N Engl J Med. 1989; 321: 1174-8). Umbilical cord blood transplantation has the advantage of increasing the number of potential donors because of the low risk of Graft versus Host disease (GVHD) and less stringent HLA matching. There is less risk of viral contamination.

The cord blood stem cells are preferably derived from human cord blood. The umbilical cord blood-derived stem cells include both mesenchymal stem cells and mesenchymal progenitor cells derived from human umbilical cord blood, and known methods (eg, Korean Patent Registration). 489,248, etc.).

The compositions of the present invention can be used to treat urinary incontinence by administration to a patient or animal. The composition of the present invention can be preferably applied to stress urinary incontinence, and can be applied to urgency incontinence, first class incontinence, reflective incontinence, and the like.

The compositions of the present invention can be administered parenterally, preferably in the form of periurethral injection injections. Compositions for parenteral administration (such as injections) of the compositions according to the invention may be infused and dispersed in pharmaceutically acceptable carriers, such as sterile purified water, buffers of about pH 7, or physiological saline, in vivo. And, if necessary, include conventional additives such as preservatives, stabilizers and the like.

The composition of the present invention can be administered to the average adult patient (about 60kg) in the dose of about 2.5 × 10 ⁸ to 6.5 × 10 ⁸ / day to the average adult patient for the purpose of treating urinary incontinence, the dose is a type of disease And depending on the state. For a typical adult patient, each unit dosage form may comprise about 4.3 ± 1.9 × 10 ⁸ cord blood derived stem cells with a suitable pharmaceutically acceptable carrier.

Hereinafter, the present invention will be described in more detail with reference to Examples, but the present invention is not limited thereto.

Example.

(1) patient selection

This study was carried out from July 2005 to December 2005 with the approval of the Institutional Ethics Committee of Gangnam Tea Hospital, Pocheon University of Medicine. Thirty-two patients with urinary incontinence received an informed consent form, and all patients had failed conservative treatment.

All patients were asked to complete a quality-of-life questionnaires and Incontinence Impact Questionnaire with pre-procedure tests, history, physical, urine, urine culture, general blood, general chemistry, A cotton swab test, a urodynamic study, and a human leukocyte antigen test (HLA typing) were performed.

The mean age of the patients in 32 patients was 51 ± 8 years old, 10 patients had anatomical incontinence, 12 patients had endogenous urethral sphincter dysfunction, 5 patients had two combinations, and 5 patients had urinary incontinence. Two of these patients had a history of previous incontinence surgery.

(2) Isolation of Stem Cells

The cord blood was to sediment the umbilical vein after normal delivery collected in CPDA-1 is 23ml umbilical cord blood collected aseptically contained bag (Green Cross, Korea), a known method (Rubinstein protocol used Processing and cryopreservation of placental / umbilical cord blood for unrelated bone marrow reconstitution Proc Natl Acad Sci USA 92: 10119-22) was used to isolate mononuclear cells from cord blood.

The same amount of saline was added to the cord blood and mixed well. Ficoll was added to a 15 ml test tube, and cord blood dilution was carefully added thereto (Piccol: cord blood dilution = 3: 4). After centrifugation at 1800 rpm for 30 minutes, the mononuclear cells present in the mononuclear cell layer on the picol were carefully extracted, and then put into a 50 ml test tube to recover the mononuclear cells. Three times the volume of physiological saline was added to the recovered mononuclear cells, and the cells were collected by centrifugation at 1800 rpm for 10 minutes. The supernatant was discarded and the remaining cell pellet was mixed well with 50 ml saline solution, and then centrifuged at 1000 rpm for 10 minutes twice. The cell pellet was suspended in 50 ml saline, and then 10 µl was taken and cell counted.

(3) injection of stem cells

The procedure of periurethral UCBSC injection was performed by one operator, and the patient was placed antibiotics before the procedure. Local anesthesia was performed and the bladder neck was visually checked using a 30 ° 26 French bladder, and the stem cells isolated in (2) were placed at 4 o'clock and 8 o'clock under the bladder at a dose of 4.3 ± 1.9 × 10 ⁸ . Urinary tract was also injected. At this time, the urethral mucosa was observed swelling with a cystoscope and injected until complete closure was achieved.

The patient was discharged on the same day after the procedure, and the patient was educated so that he could be immediately visited and treated for abnormal symptoms. He performed physical examination and liver function tests every week until the month after the procedure. To evaluate the results of the procedure, the urination diary and patient satisfaction were compared one month and three months after the procedure, and objective urinary incontinence was confirmed by urethral dynamics test three months after the procedure.

(4) results

The average duration of periurethral UCBSC injection was 15 minutes, and 6 patients (18.8%) had leaked into the injected urethral mucosa at the beginning of the study. There were two. Postoperative urination disorders, urine congestion, urethral infection, and GVHD (Graft versus Host Disease) did not occur in any of the cases (see Table 1).

Symptom Number Procedure failure Injection fluid leak 6 (18.8%) Pain caused by pain 2 (6.3%) Postoperative Symptoms Urination disorder 0 Urine stagnation 0 Urethral infection 0 GVHD 0

The mean follow-up was 5.1 months (range 3-8 months). Three months after the operation, three patients were dropped out and 18 patients (56.3%) showed improvement of symptoms. Four patients (12.5%) had no improvement, and in the satisfaction survey conducted three months after the operation, ten patients were further departed, but 15 patients (49.6) still showed improvement of more than 50%. Reference).

After one month 3 months later Improvement of symptoms ≥ 50% 18 (56.3%) 15 (46.9%) > 50% 7 (21.9%) 1 (3.1%) No change (≤10%) 4 (12.5%) 3 (9.4%) Breakaway 3 (9.4%) 13 (40.6%) Sum 32 (100%) 32 (100%)

In addition, it was confirmed that the by the urodynamic testing from preoperative stress urinary incontinence five people due to the intrinsic urethral sphincter dysfunction performed 3 months after surgery, the maximum urethral closure pressure from all five people improve more than 30 cmH ₂ O (Table 3 Reference).

Preoperative urodynamic examination Preoperative urodynamic examination <30 cmH ₂ O <30 cmH ₂ O ≥ 30 cmH ₂ O Follow-up 5 0 5 (100%) Breakaway 12 Sum 17

As described above, the urinary incontinence treatment composition of the present invention, when injected into the urethra, can be treated in a very short time hospital treatment, it can be performed without complications associated with surgery. So similar regardless of the type of stress urinary incontinence showed a subjective improvement of 78.2% at 1 month after surgery, maximum urethral closure pressure (MUCP) is 30 cmH ₂ O or less was 5 people both in the operative urodynamic testing was performed after 3 months 30 Improved results were observed above cmH ₂ O.

Urinary incontinence treatment composition comprising umbilical cord blood-derived stem cells according to the present invention can effectively treat urinary incontinence when injected in vivo, especially when injected into the urethral infusion, minimizing postoperative complications compared to conventional surgery Urinary incontinence can be treated by a very short period of time.

Claims (3)

Umbilical cord blood stem cells (umbilical cord blood stem cell) and a pharmaceutically acceptable carrier, a composition for treating incontinence. The composition of claim 1, wherein said urinary incontinence is a stress urinary incontinence. The composition of claim 1 or 2, wherein the formulation of the composition is in the form of a periurethral injectable.